Team:KIT-Kyoto/Notebook/YJL

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Revision as of 06:06, 27 September 2013

YJL068C

13th August

We designed the PCR primers to amplify YJL068C gene.

19^th August

1. Make Primer mix

We added 102.2µL and 100.3µL H₂O to the forward and reverse primers, respectively, to make 100 pmol/µL solutions.

We obtained 10µL of each primer solution and added 80µL H₂O to make the 10 pmol/µL primer mix.

2. PCR

We performed PCR to amplify the YJL068C gene.

Buffer	50µL
dNTP	20µL
Primer mix	1µL
Genome DNA	0.5µL
KOD-FX	2µL
H₂O	26.5µL

(Annealing : 52 ˚C)

We detected two DNA bands around 1400bp and 3000bp.

(The correct band is 1400bp.)

We changed annealing temperature to 54°C or 57 ˚C and tried PCR again.

We detected a correct size DNA band around 1400bp.

The clear DNA band was amplified at 54 ˚C of annealing temperature.

We performed PCR to amplify YJL068C gene at 54°C of annealing temp (total volume was 500µL).

20^th August

1

We performed PCR to check whether the primers can work or not.

(Annealing temperature: 55°C)

We detected a DNA band of the correct size around 900bp.

2

We purified the YJL068C gene (PCR was performed on 8/19).

We digested YJL068C with BanII to check whether PCR succeeded or not.

We ligated YJL068C and pSB1C3.

YJL068C	24µL
M Buffer	3µL
BSA	1µL
XbaI	1µL
SpeI	1µL

YJL068C	6µL
H Buff.	2µL
H2O	11µL
BanII	1µL

We digested psb1c3 with XbaI and SpeI at 37°C for 90 minutes.

pSB1C3	49µL
M Buffer	5.78µL
BSA	1µL
XbaI	1µL
SpeI	1µL

We added 1µL BAP to dephosphorylate the pcb1c3 gene.

We electrophoresed the YJL068C gene and psb1c3 (digested for ligation) in 1% Blue gel.

We saw the clear band of YJL068C, and not so clear band of psb1c3.

They were extracted from the Blue gel.

We prepared the spare the spare in case of failure (made stocks of purified YJL068C and psb1c3).

We purified YJL068C and psb1c3 (these were in one tube) and ligated them.

We transformed it all (20µL) into E.coli cells.

21th August

1

We checked the YJL068C gene (digested with Ban2 8/20) by electrophoresing.

We could see correct band (succeeded PCR).

2

We checked of the insert by colony cracking (transformed 8/20).

We couldn’t see clear bands, but cultured furthermore.

3.

We made the spare in case of failure (prepared 8/20).

We purified the YJL068C gene (digested 8/20).

We digested psb1c3 with XbaI and SpeI (90 minutes at 37°C), and added 1µL BAP to it.

pSB1C3	96µL
Buffer	11µL
BSA	1µL
XbaI	1µL
SpeI	1µL

We electrophoresed psb1c3 it in 1% Blue gel.

It was extracted from the Blue gel.

YJL068C	5µL
pSB1C3	5µL
Ligation Mix	10µL

We added 5µL H₂O to dried YJL068C gene and psb1c3 each.

We ligated them and transformed it into E.coli cells.

22th August

1.

We did miniprep to purify DNA.

We digested YJL068C in pSB1C3 with PstI and EcorI, and incubated at 37°C for 90 minutes.

Plasmid DNA	10µL
H Buffer	2µL
H₂O	6µL
EcoRI	1µL
PstI	1µL

We applied them to 1% agarose gel electrophoresis and checked them.

5 samples showed the correct size.

2.

We digested pSB1C3 with BsiW1.

pSB1C3	10µL
NEB Buffer 3	2µL
BsiWI	1µL
H₂O	7µL

23th August

1.

We applied YJL068C in pSB1C3 (prepared on 8/22) to 1% agarose gel electrophoresis.

We detected the correct size of DNA.

Each transformant was cultured in LB liquid medium containing Chloramphenicol.

Plasmid DNA was purified by miniprep.

24th August

We added 29.5µL of H₂O to purified YJL068C.

We digested YJL068C with BanII and checked.

YJL068C	5µL
H Buff.	2µL
H₂O	12µL
BanII	1µL

We detected the correct size of DNA band.

We digested YJL068C with SpeI and BamHI.

YJL068C	24.5µL
Buffer	3µL
BSA	0.5µL
SpeI	1µL
BamHI	1µL

We digested pET-15b with XbaI and BamHI and dephosphorylated with 1uL BAP.

pET-15b	87µL
Buffer	10µL
BSA	1µL
XbaI	1µL
BamHI	1µL

We extracted DNA from the Blue gel.

We added 10µL H₂O to dried YJL068C gene and pET-15b.

We ligated them and transformed it into E.coli cells.

25th August

We cultured them furthermore.

26th August

We checked the insertion of DNA by colony cracking.

We cultured each transformant in LB liquid medium containing ampicillin.

We added 20µL of IPTG to them and incubated.

Cell free extract was prepared and applied to SDS-PAGE to detecte ATF2 protein.

29th August

Cell free extract was prepared with FastBreak Cell Lysis Reagent and applied to SDS-PAGE.

30th August

We performed PCR to amplify YJL068C.

(Annealing temperature: 55°C)

31th August

We purified the YJL068C (performed PCR at 8/30).

We digested YJL068C with BanII to check whether PCR succeeded or not.

We ligated YJL068C and pSB1C3.

We digested YJL068C with XhoI and NdeI for ligation.

We purified pET-15b and digested with XhoI and NdeI.

YJL068C	24µL
Buffer	3µL
BSA	1µL
XhoI	1µL
NdeI	1µL

pET-15b	33µL
Buffer	4µL
BSA	1µL
XhoI	1µL
NdeI	1µL

DNA samples were applied to 1% Blue gel electrophoresis and then purified.

We added 10µL H₂O to dried YJL068C gene and pET-15b.

We ligated them and transformed it into E.coli cells.

September 1^st

Picked 32 colonies of YJL068C transformants.

September 4th

Digested YJL068C and pET-15b overnight.

Applied pET-15b and ATF1 to the blue gel electrophoresis.

Isolated and purified it.

September 5^th

Purified PCR products and dissolved 24µL of H₂O.

Purified pET-15b.

Digested YJL068C and pET-15b with XhoI and NdeI overnight.

YJL068C	24µL
Buffer	3µL
BSA	1µL
XhoI	1µL
NdeI	1µL

pET-15b	33µL
Buffer	4µL
BSA	1µL
XhoI	1µL
NdeI	1µL

September 6th

1.

Applied YJL068C and pET-15b(prepared on September 5th) to the blue gel electrophoresis.

Isolated and purified it.

Ligation of pET-15b with YJL 068C (prep).

YJL068C	10µL
pET-15b	10µL
Ligation Mix	10µL

Transformed YJL068C into pET-15b E. coli cells.

2.

Purified PCR products and pET-15b were dissolved in 24.5µL of H₂O.

YJL068C and pET-15b were digested with XhoⅠand NdeⅠ(overnight).

YJL068C	24.5µL
Buffer	3µL
BSA	0.5µL
XhoI	1µL
NdeI	1µL

pET-15b	24.5µL
Buffer	3µL
BSA	0.5µL
XhoI	1µL
NdeI	1µL

Applied YJL068C and pET-15b(prepared on September 5th) to the blue gel electrophoresis.

Isolated and purified it.

Ligation of pET-15b with YJL (prep).

Transformed YJL068C into pET-15b E. coli cells.

September 7^th

Picked 48 colonies of YJL068C transformants.

Checked the colonies by colony cracking.

Picked up the appropriate colonies and cultured in 3mL LB medium with ampicillin.

September 8^th

Plasmid DNA was prepared by miniprep and digested with XhoI/NdeI.

(1)

Plasmid DNA	15.5µL
Buffer	2µL
BSA	0.5µL
XhoI	1µL
NdeI	1µL

(2)

BsiWI	1µL
Buffer	2µL
Plasmid DNA	17µL

Electrophoresed in 1% agarose gel.

Not succeeded.

September 15^th

Transformed YJL068C into pET-15b E. coli cells.

September 16^th

Picked 48 colonies of YJL068C transformants.

September 17th

Checked the colonies by colony cracking.

Picked up the colonies and cultured in 3mL LB medium with ampicillin.

September 18^th

Miniprepped plasmid DNA(YJL068C into pET-15b) and digested it.

Electrophoresed in 1% agarose gel.

Not succeeded.

September 19^th

YJL068C and pET-15b were digested with XhoI and BlpI .

YJL068C	24.5µL
NEB Buffer 4	3µL
BSA	0.5µL
XhoI	1µL
BlpI	1µL

pET-15b	24.5µL
NEB Buffer 4	3µL
BSA	0.5µL
XhoI	1µL
BlpI	1µL

Transformed YJL068C into pET-15b E. coli cells.

September 20^th

Picked 64 colonies of YJL068C transformants.

September 21st

Checked the colonies by colony cracking.

Picked up the appropriate colonies and cultured in the LB in 3mL ampicillin(+) medium.

September 22^nd

Miniprepped plasmid DNA(YJL068C into pET-15b) and digested it.

Plasmid DNA	16µL
XhoI	1µL
BlpI	1µL
NEB Buffer 4	2µL

Electrophoresed in 1% agarose gel.

We could not get any YJL068C into pET-15b E. coli cells.

@@ Line 16: / Line 16: @@
 <h2>YJL068C</h2>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>13th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-designed the PCR primers to amplify</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>
-YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt"> gene.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>19</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP><FONT SIZE=4><B>
-August</B></FONT></SPAN></FONT></P>
-<OL>
-	<LI><P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Make
-	Primer mix</FONT></SPAN></FONT></P>
-</OL>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-added 102.2 uL and 100.3 uL H2O to the forward and reverse primers,
-respectively, to make 100 pmol/microL solutions.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-obtained 10 uL of each primer solution and added 80 uL H2O to make
-the 10 pmol/microL primer mix.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<OL START=2>
-	<LI><P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">PCR</FONT></SPAN></FONT></P>
-</OL>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-performed PCR to amplify the </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">50ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">dNTP</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">20ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Primer
-			mix</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Genome
-			DNA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">0.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">KOD-FX</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=147>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">26.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"> <FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">(Annealing
-: 52&deg;C)</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-detected two DNA bands around 1400bp and 3000bp.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">(The
-correct band is 1400bp.)</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-changed annealing temperature to 54&deg;C or 57&deg;C and tried PCR
-again.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-detected a correct size DNA band around 1400bp.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">The
-clear DNA band was amplified at 54&#730;C of annealing temperature.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-performed PCR to amplify </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene at 54&deg;C of annealing temp (total volume was 500 microL).</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>20</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP><FONT SIZE=4><B>
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-performed PCR to check whether the primers can work or not.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">(Annealing
-temperature: 55&deg;C)</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-detected a DNA band of the correct size around 900bp.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-purified the </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene (PCR was performed on 8/19).</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-with </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>Ban</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">2
-to check whether PCR succeeded or not.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-ligated </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-and pSB1C3.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">24ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">M
-			Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xba1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Spe1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<TABLE WIDTH=145 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">6ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H
-			Buff.</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">11ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BanⅡ</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested psb1c3 with XbaI and SpeI at 37&deg;C for 90 minutes.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pSB1C3</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">49ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">M
-			Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">5.78ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xba1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Spe1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-added 1uL BAP to dephosphorylate the </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>pcb1c3</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-electrophoresed the </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I> </I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">and
-psb1c3 (digested for ligation) in 1% Blue gel.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-saw the clear band of YJL068C, and not so clear band of psb1c3.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">They
-were extracted from the Blue gel.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-prepared the spare the spare in case of failure (made stocks of
-purified YJL068C and psb1c3).</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-purified YJL068C and psb1c3 (these were in one tube) and ligated
-them.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-transformed it all (20 microL) into </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>E.coli</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-cells.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm; page-break-before: always"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>21th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-checked the </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene (digested with </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>Ban</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">2
-/20) by electrophoresing.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-could see correct band (succeeded PCR).</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-checked of the insert by colony cracking (transformed 8/20).</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-couldn&rsquo;t see clear bands, but cultured furthermore.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-made the spare in case of failure (prepared 8/20).</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-purified the </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene (digested 8/20).</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested psb1c3 with </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>Xba</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">I
-and </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>Spe</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">I
-(90 minutes at 37&deg;C), and added 1uL BAP to it.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pSB1C3</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">96ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">11ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xba1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Spe1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-electrophoresed psb1c3 it in 1% Blue gel.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">It
-was extracted from the Blue gel.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=170 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=99>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=99>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pSB1C3</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=99>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligation
-			Mix</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">10ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-added 5 microL H2O to dried </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene and psb1c3 each.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-ligated them and transformed it into </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>E.coli
-</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">cells.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>22th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-did miniprep to purify DNA.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-in pSB1C3 with Pst1 and EcoR1, and incubated at 37&deg;C for 90
-minutes.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=176 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Plasmid
-			DNA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">10ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H
-			Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">6ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">EcoR1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Pst1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-applied them to 1% agarose gel electrophoresis and checked them.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">5
-samples showed the correct size.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested pSB1C3 with BsiW1.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=163 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=92>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pSB1C3</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">10ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=92>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3
-			NEB Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=92>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BsiW1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=92>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">7ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>23th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-applied </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-in pSB1C3 (prepared on 8/22) to 1% agarose gel electrophoresis.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-detected the correct size of DNA.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Each
-transformant was cultured in LB liquid medium containing
-Chloramphenicol.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Plasmid
-DNA was purified by miniprep.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>24th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-added 29.5uL of H2O to purified YJL068C.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-with BanII and checked.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=145 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H
-			Buff.</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">12ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BanⅡ</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-detected the correct size of DNA band.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I> YJL068C </I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">with
-SpeI and BamHI.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">24.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">0.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Spe1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BamH1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested pET-15b with XbaI and BamHI and dephosphorylated with 1uL
-BAP.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">87ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">10ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xba1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BamH1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-extracted DNA from the Blue gel.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-added 10 uL H2O to dried </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene and pET-15b.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-ligated them and transformed it into </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>E.coli
-</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">cells.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>25th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-cultured them furthermore.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>26th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-checked the insertion of DNA by colony cracking.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-cultured each transformant in LB liquid medium containing ampicillin.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-added 20ul of IPTG to them and incubated.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Cell
-free extract was prepared and applied to SDS-PAGE to detecte ATF2
-protein.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>29th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Cell
-free extract was prepared with FastBreak Cell Lysis Reagent and
-applied to SDS-PAGE.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>30th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-performed PCR to amplify </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">(Annealing
-temperature: 55&deg;C)</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>31th
-August</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-purified the </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-(performed PCR at 8/30).</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-with </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>Ban</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">II
-to check whether PCR succeeded or not.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-ligated </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-and pSB1C3.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-digested </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-with </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>Xho</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">I
-and </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>Nde</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">I
-for ligation.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-purified pET-15b and digested with </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>Xho</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">I
-and </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>Nde</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">I.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">24ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Nde1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">33ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">4ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Nde1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">DNA
-samples were applied to 1% Blue gel electrophoresis and then
-purified.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-added 10 uL H2O to dried </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-gene and pET-15b.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-ligated them and transformed it into </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>E.coli
-</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">cells.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>st</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
-colonies of </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-transformants.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-th</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Digested
-</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-and pET-15b overnight.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Applied
-pET-15b and </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-to the blue gel electrophoresis.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Isolated
-and purified it.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Purified
-PCR products and dissolved 24ul of H2O.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Purified
-pET-15b.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Digested
-</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-and pET-15b with XhoⅠand NdeⅠ overnight.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">24ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Nde1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">33ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">4ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Nde1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm; page-break-before: always"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-th</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Applied
-</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-and pET-15b(prepared on September 5th) to the blue gel
-electrophoresis.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Isolated
-and purified it.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligation
-of pET-15b with </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-(prep).</FONT></SPAN></FONT></P>
-<TABLE WIDTH=170 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=99>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">10ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=99>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">10ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=99>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligation
-			Mix</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">10ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Transformed
-</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-into pET-15b </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>E. coli</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-cells.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Purified
-PCR products and pET-15b were dissolved in 24.5ul of H2O.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-and pET-15b were digested with XhoⅠand NdeⅠ(overnight).</FONT></SPAN></FONT></P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">24.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">0.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Nde1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<TABLE WIDTH=146 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">24.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">0.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=75>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Nde1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Applied
-YJL068C</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-and pET-15b(prepared on September 5th) to the blue gel
-electrophoresis.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Isolated
-and purified it.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligation
-of pET-15b with YJL068C (prep).</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Transformed
-YJL068C</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-into pET-15b E. coli cells.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
-colonies of </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C
-</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">transformants.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Checked
-the colonies by colony cracking.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
-up the appropriate colonies and cultured in 3ml LB medium with
-ampicillin.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm; page-break-before: always"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Plasmid
-DNA was prepared by miniprep and digested with XhoI/NdeI.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">(1)</FONT></SPAN></FONT></P>
-<TABLE WIDTH=176 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Plasmid
-			DNA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">15.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">0.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Nde1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">(2)</FONT></SPAN></FONT></P>
-<TABLE WIDTH=176 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BsiW1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Plasmid
-			DNA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">17ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Electrophoresed
-in 1% agarose gel.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Not
-succeeded.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Transformed
-</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-into pET-15b </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>E. coli</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-cells.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
-colonies of </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C
-</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">transformants.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-th</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Checked
-the colonies by colony cracking.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
-up the colonies and cultured in 3ml LB medium with ampicillin.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Miniprepped
-plasmid DNA(</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-into pET-15b) and digested it.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Electrophoresed
-in 1% agarose gel.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Not
-succeeded.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm; page-break-before: always"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-and pET-15b were digested with XhoⅠand BlpⅠ .</FONT></SPAN></FONT></P>
-<TABLE WIDTH=176 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">24.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">NEB
-Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">0.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Blp1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<TABLE WIDTH=176 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">24.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">NEB
-Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">0.5ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=104>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Blp1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=42>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Transformed
-</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-into pET-15b </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>E. coli</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-cells.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
-colonies of YJL068C transformants.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-st</B></FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Checked
-the colonies by colony cracking.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
-up the appropriate colonies and cultured in the LB in 3ml
-ampicillin(+) medium.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
-</B></FONT><SUP><FONT SIZE=4><B>nd</B></FONT></SUP></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Miniprepped
-plasmid DNA(</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-into pET-15b) and digested it.</FONT></SPAN></FONT></P>
-<TABLE WIDTH=176 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Plasmid
-			DNA</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">16ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Blp1</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-	<TR VALIGN=TOP>
-		<TD WIDTH=105>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">NEB
-Buffer</FONT></SPAN></FONT></P>
-		</TD>
-		<TD WIDTH=41>
-			<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
-		</TD>
-	</TR>
-</TABLE>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Electrophoresed
-in 1% agarose gel.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
-could not get any </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>YJL068C</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-into pET-15b </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>E. coli</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
-cells.</FONT></SPAN></FONT></P>
-<P STYLE="margin-bottom: 0cm"><BR>
-</P>
+<div class=WordSection1 style='layout-grid:18.0pt'>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>13th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We designed the
+PCR primers to amplify<i> YJL068C</i> gene.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>19<sup>th</sup>
+August</span></b></p>
+<p class=MsoNormal style='margin-left:18.0pt;text-indent:-18.0pt'><span
+lang=EN-US style='font-size:11.0pt'>1.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+</span></span><span lang=EN-US style='font-size:11.0pt'>Make Primer mix</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We added 102.2</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L and 100.3</span><span lang=EN-US style='font-size:11.0pt'>&micro;</span><span
+lang=EN-US style='font-size:11.0pt'>L H<sub>2</sub>O to the forward and reverse
+primers, respectively, to make 100 pmol/</span><span lang=EN-US
+style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:11.0pt'>L
+solutions.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We obtained 10</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L of each primer solution and added 80</span><span lang=EN-US
+style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:11.0pt'>L H<sub>2</sub>O
+to make the 10 pmol/</span><span lang=EN-US style='font-size:11.0pt'>&micro;</span><span
+lang=EN-US style='font-size:11.0pt'>L primer mix.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal style='margin-left:18.0pt;text-indent:-18.0pt'><span
+lang=EN-US style='font-size:11.0pt'>2.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+</span></span><span lang=EN-US style='font-size:11.0pt'>PCR</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR
+to amplify the <i>YJL068C</i> gene.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>50</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>dNTP</span></p>
+  </td>
+  <td width=162 valign=top style='width:121.45pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Primer mix</span></p>
+  </td>
+  <td width=162 valign=top style='width:121.45pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Genome DNA</span></p>
+  </td>
+  <td width=162 valign=top style='width:121.45pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>KOD-FX</span></p>
+  </td>
+  <td width=162 valign=top style='width:121.45pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H<sub>2</sub>O</span></p>
+  </td>
+  <td width=162 valign=top style='width:121.45pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;(Annealing :
+</span><span lang=EN-US> </span><span lang=EN-US style='font-size:11.0pt'>&#730;C)</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We detected two
+DNA bands around 1400bp and 3000bp.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>(The correct band
+is 1400bp.)</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We changed
+annealing temperature to 54°C or 57</span><span lang=EN-US> </span><span
+lang=EN-US style='font-size:11.0pt'>&#730;C and tried PCR again.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We detected a
+correct size DNA band around 1400bp.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>The clear DNA band
+was amplified at 54</span><span lang=EN-US> </span><span lang=EN-US
+style='font-size:11.0pt'>&#730;C of annealing temperature.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR
+to amplify <i>YJL068C</i> gene at 54°C of annealing temp (total volume was 500</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L).</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>20<sup>th</sup>
+August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR
+to check whether the primers can work or not.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>(Annealing temperature:
+°C)</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We detected a DNA
+band of the correct size around 900bp.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We purified the <i>YJL068C</i>
+gene (PCR was performed on 8/19).</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested <i>YJL068C</i>
+with <i>Ban</i>II to check whether PCR succeeded or not.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We ligated <i>YJL068C</i>
+and pSB1C3.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>24</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>M Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XbaI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>SpeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=56 valign=top style='width:41.65pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>6</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H Buff.</span></p>
+  </td>
+  <td width=56 valign=top style='width:41.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H2O</span></p>
+  </td>
+  <td width=56 valign=top style='width:41.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>11</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BanII</span></p>
+  </td>
+  <td width=56 valign=top style='width:41.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested psb1c3
+with XbaI and SpeI at 37°C for 90 minutes.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pSB1C3</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>49</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>M Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>5.78</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xba</span></i><span
+  lang=EN-US style='font-size:11.0pt'>I</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>SpeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We added 1</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L BAP to dephosphorylate the <i>pcb1c3</i> gene.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We electrophoresed
+the <i>YJL068C</i> gene<i> </i>and psb1c3 (digested for ligation) in 1% Blue gel.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We saw the clear
+band of YJL068C, and not so clear band of psb1c3.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>They were
+extracted from the Blue gel.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We prepared the spare
+the spare in case of failure (made stocks of purified YJL068C and psb1c3).</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We purified
+YJL068C and psb1c3 (these were in one tube) and ligated them.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We transformed it
+all (20</span><span lang=EN-US style='font-size:11.0pt'>&micro;</span><span
+lang=EN-US style='font-size:11.0pt'>L) into <i>E.coli</i> cells.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<span lang=EN-US style='font-size:11.0pt;font-family:"Century","serif"'><br
+clear=all style='page-break-before:always'>
+</span>
+<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
+style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>21th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We checked the <i>YJL068C</i>
+gene (digested with <i>Ban</i>2 8/20) by electrophoresing.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We could see
+correct band (succeeded PCR).</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We checked of the
+insert by colony cracking (transformed 8/20).</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We couldn’t see
+clear bands, but cultured furthermore.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We made the spare
+in case of failure (prepared 8/20).</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We purified the <i>YJL068C</i>
+gene (digested 8/20).</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested psb1c3
+with <i>Xba</i>I and <i>Spe</i>I (90 minutes at 37°C), and added 1</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L BAP to it.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pSB1C3</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>96</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>11</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xba</span></i><span
+  lang=EN-US style='font-size:11.0pt'>I</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>SpeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We electrophoresed
+psb1c3 it in 1% Blue gel.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>It was extracted
+from the Blue gel.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=114 valign=top style='width:85.45pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=114 valign=top style='width:85.45pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pSB1C3</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=114 valign=top style='width:85.45pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligation Mix</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We added 5</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L H<sub>2</sub>O to dried <i>YJL068C</i> gene and psb1c3 each.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We ligated them
+and transformed it into <i>E.coli </i>cells.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>22th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We did miniprep to
+purify DNA.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested <i>YJL068C</i>
+in pSB1C3 with PstI and EcorI, and incubated at 37°C for 90 minutes.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Plasmid DNA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H<sub>2</sub>O</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>6</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>EcoRI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>PstI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied them to
+% agarose gel electrophoresis and checked them.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>5 samples showed the
+correct size.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested pSB1C3
+with BsiW1.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=107 valign=top style='width:80.1pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pSB1C3</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=107 valign=top style='width:80.1pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NEB Buffer 3</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=107 valign=top style='width:80.1pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BsiWI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=107 valign=top style='width:80.1pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H<sub>2</sub>O</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>7</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>23th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied <i>YJL068C</i>
+in pSB1C3 (prepared on 8/22) to 1% agarose gel electrophoresis.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We detected the
+correct size of DNA.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Each transformant
+was cultured in LB liquid medium containing Chloramphenicol.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Plasmid DNA was
+purified by miniprep.</span></p>
+<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>24th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We added 29.5</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L of H<sub>2</sub>O to purified YJL068C.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested <i>YJL068C</i>
+with BanII and checked.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=56 valign=top style='width:41.65pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H Buff.</span></p>
+  </td>
+  <td width=56 valign=top style='width:41.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H<sub>2</sub>O</span></p>
+  </td>
+  <td width=56 valign=top style='width:41.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>12</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BanII</span></p>
+  </td>
+  <td width=56 valign=top style='width:41.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We detected the
+correct size of DNA band.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested<i>
+YJL068C </i>with SpeI and BamHI.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>24.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>SpeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BamHI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested
+pET-15b with XbaI and BamHI and dephosphorylated with 1uL BAP.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>87</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XbaI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BamHI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We extracted DNA
+from the Blue gel.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We added 10</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L H<sub>2</sub>O to dried <i>YJL068C</i> gene and pET-15b.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We ligated them
+and transformed it into <i>E.coli </i>cells.</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>25th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We cultured them
+furthermore.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>26th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We checked the
+insertion of DNA by colony cracking.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We cultured each
+transformant in LB liquid medium containing ampicillin.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We added 20</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L of IPTG to them and incubated.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Cell free extract
+was prepared and applied to SDS-PAGE to detecte ATF2 protein.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>29th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Cell free extract
+was prepared with FastBreak Cell Lysis Reagent and applied to SDS-PAGE.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>30th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR to
+amplify <i>YJL068C</i>.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>(Annealing temperature:
+°C)</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>31th August</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We purified the <i>YJL068C</i>
+(performed PCR at 8/30).</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested <i>YJL068C</i>
+with <i>Ban</i>II to check whether PCR succeeded or not.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We ligated <i>YJL068C</i>
+and pSB1C3.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested <i>YJL068C</i>
+with <i>Xho</i>I and <i>Nde</i>I for ligation.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We purified
+pET-15b and digested with <i>Xho</i>I and <i>Nde</i>I.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>24</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NdeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>33</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>4</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NdeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>DNA samples were
+applied to 1% Blue gel electrophoresis and then purified.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We added 10</span><span
+lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L H<sub>2</sub>O to dried <i>YJL068C</i> gene and pET-15b.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We ligated them
+and transformed it into <i>E.coli </i>cells.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 1<sup>st</sup></span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked 32 colonies
+of <i>YJL068C</i> transformants.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 4th</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested <i>YJL068C</i>
+and pET-15b overnight.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied pET-15b
+and <i>ATF1</i> to the blue gel electrophoresis.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Isolated and purified
+it.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 5<sup>th</sup></span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified PCR
+products and dissolved 24&micro;L of H<sub>2</sub>O.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified pET-15b.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested <i>YJL068C</i>
+and pET-15b with XhoI and NdeI overnight.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>24</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NdeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>33</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>4</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NdeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<span lang=EN-US style='font-size:11.0pt;font-family:"Century","serif"'><br
+clear=all style='page-break-before:always'>
+</span>
+<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
+style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 6th</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied <i>YJL068C</i>
+and pET-15b(prepared on September 5th) to the blue gel electrophoresis.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Isolated and purified
+it.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligation of
+pET-15b with <i>YJL</i> <i>068C</i> (prep).</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=114 valign=top style='width:85.45pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=114 valign=top style='width:85.45pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=114 valign=top style='width:85.45pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligation Mix</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Transformed <i>YJL068C</i>
+into pET-15b <i>E. coli</i> cells.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified PCR
+products and pET-15b were dissolved in 24.5</span><span lang=EN-US
+style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:11.0pt'>L of
+H<sub>2</sub>O.</span></p>
+<p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i><span
+lang=EN-US style='font-size:11.0pt'> and pET-15b were digested with Xho</span><span
+style='font-size:11.0pt;font-family:"ＭＳ 明朝","serif"'>Ⅰ</span><span lang=EN-US
+style='font-size:11.0pt'>and Nde</span><span style='font-size:11.0pt;
+font-family:"ＭＳ 明朝","serif"'>Ⅰ</span><span lang=EN-US style='font-size:11.0pt'>(overnight).</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>24.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NdeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>24.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=90 valign=top style='width:67.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NdeI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied YJL<i>068C</i>
+and pET-15b(prepared on September 5th) to the blue gel electrophoresis.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Isolated and
+purified it.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligation of
+pET-15b with YJL (prep).</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Transformed YJL<i>068C</i>
+into pET-15b E. coli cells.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 7<sup>th</sup></span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked 48 colonies
+of <i>YJL068C </i>transformants.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Checked the
+colonies by colony cracking.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked up the
+appropriate colonies and cultured in 3mL LB medium with ampicillin.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<span lang=EN-US style='font-size:11.0pt;font-family:"Century","serif"'><br
+clear=all style='page-break-before:always'>
+</span>
+<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
+style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 8<sup>th</sup></span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Plasmid DNA was
+prepared by miniprep and digested with XhoI/NdeI.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>(1)</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Plasmid DNA</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>15.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NdeI</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>(2)</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BsiWI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Plasmid DNA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>17</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Electrophoresed in
+% agarose gel.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Not succeeded.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 15<sup>th</sup></span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Transformed <i>YJL068C</i>
+into pET-15b <i>E. coli</i> cells.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 16<sup>th</sup></span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked 48 colonies
+of <i>YJL068C </i>transformants.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 17th</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Checked the
+colonies by colony cracking.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked up the
+colonies and cultured in 3mL LB medium with ampicillin.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 18<sup>th</sup></span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Miniprepped
+plasmid DNA(<i>YJL068C</i> into pET-15b) and digested it.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Electrophoresed in
+% agarose gel.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Not succeeded.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<span lang=EN-US style='font-size:11.0pt;font-family:"Century","serif"'><br
+clear=all style='page-break-before:always'>
+</span>
+<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
+style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 19<sup>th</sup></span></b></p>
+<p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i><span
+lang=EN-US style='font-size:11.0pt'> and pET-15b were digested with XhoI and
+BlpI .</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>YJL068C</span></i></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>24.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NEB Buffer 4</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BlpI</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>24.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NEB Buffer 4</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=119 valign=top style='width:89.3pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BlpI</span></p>
+  </td>
+  <td width=57 valign=top style='width:42.4pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Transformed <i>YJL068C</i>
+into pET-15b <i>E. coli</i> cells.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 20<sup>th</sup></span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked 64 colonies
+of YJL068C transformants.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 21st</span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Checked the
+colonies by colony cracking.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked up the
+appropriate colonies and cultured in the LB in 3mL ampicillin(+) medium.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 22<sup>nd</sup></span></b></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Miniprepped
+plasmid DNA(<i>YJL068C</i> into pET-15b) and digested it.</span></p>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none'>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Plasmid DNA</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border:solid windowtext 1.0pt;
+  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>16</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>XhoI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BlpI</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+ <tr>
+  <td width=120 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
+  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NEB Buffer 4</span></p>
+  </td>
+  <td width=56 valign=top style='width:42.25pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0mm 5.4pt 0mm 5.4pt'>
+  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span
+  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
+.0pt'>L</span></p>
+  </td>
+ </tr>
+</table>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Electrophoresed in
+% agarose gel.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We could not get
+any <i>YJL068C</i> into pET-15b <i>E. coli</i> cells.</span></p>
+<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
+</div>
 </div>
 </div>