Team:KIT-Kyoto/Notebook/ATF1/september

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<div class="document">
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<ul class="nav metro-nav-list">
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{{Template:KIT-Kyoto/menu_note}}
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  <li class="nav-header">LabNote</li>
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      <a href="#ATF1">ATF1</a>
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      <ul class="nav">
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          <li><a href="https://2013.igem.org/Team:KIT-Kyoto/Notebook/ATF1/may">May</a></li>
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          <li><a href="">june</a></li>
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          <li><a href="https://2013.igem.org/Team:KIT-Kyoto/Notebook/ATF1/july">july</a></li>
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          <li><a href="https://2013.igem.org/Team:KIT-Kyoto/Notebook/ATF1/august">August</a></li>
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          <li><a href="https://2013.igem.org/Team:KIT-Kyoto/Notebook/ATF1/september">September</a></li>
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      </ul>
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      <a href="#https://2013.igem.org/Team:KIT-Kyoto/Notebook/ATF2">ATF2</a>
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      <a href="https://2013.igem.org/Team:KIT-Kyoto/Notebook/YJL">YJL068C</a>
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      <a href="https://2013.igem.org/Team:KIT-Kyoto/Notebook/growcurves">Growth Curves</a>
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<h2>ATF1</h2>
<h2>ATF1</h2>
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 1<sup>st</sup></span></b></p>
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
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<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked 32 colonies
-
1</B></FONT><SUP><FONT SIZE=4><B>st</B></FONT></SUP></SPAN></FONT></P>
+
of <i>ATF1 </i>transformants.</span></p>
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
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32 colonies of </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1
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</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">transformants.</FONT></SPAN></FONT></P>
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<P STYLE="margin-bottom: 0cm"><BR>
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</P>
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
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3</B></FONT><SUP><FONT SIZE=4><B>rd</B></FONT></SUP></SPAN></FONT></P>
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Checked
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the colonies by colony cracking.</FONT></SPAN></FONT></P>
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">No
+
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appropriate plasmid DNA was detected.</FONT></SPAN></FONT></P>
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Digested
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</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
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and pET-15b with  at 37℃ overnight.</FONT></SPAN></FONT></P>
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-
<TABLE WIDTH=166 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
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-
<TR VALIGN=TOP>
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<TD WIDTH=96>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">ATF1</FONT></SPAN></FONT></P>
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-
</TD>
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-
<TD WIDTH=40>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">24ul</FONT></SPAN></FONT></P>
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-
</TD>
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</TR>
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<TR VALIGN=TOP>
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<TD WIDTH=96>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
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-
</TD>
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<TD WIDTH=40>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
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-
</TD>
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-
</TR>
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<TR VALIGN=TOP>
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-
<TD WIDTH=96>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu11021</FONT></SPAN></FONT></P>
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-
</TD>
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<TD WIDTH=40>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
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<TR VALIGN=TOP>
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<TD WIDTH=96>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
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-
</TD>
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<TD WIDTH=40>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
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-
</TD>
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</TR>
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<TR VALIGN=TOP>
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<TD WIDTH=96>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2NEB
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-
Buffer</FONT></SPAN></FONT></P>
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</TD>
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<TD WIDTH=40>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
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-
</TD>
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-
</TR>
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</TABLE>
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-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
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-
<TABLE WIDTH=166 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
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-
<TR VALIGN=TOP>
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-
<TD WIDTH=96>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
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<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=96>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<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=96>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu11021</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<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=96>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<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=96>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2NEB
+
-
Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">4ul</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>September
+
-
4</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">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">No
+
-
pET-15b was detected.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Extracted
+
-
from Blue gel and purified</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>
+
-
ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">.</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
+
-
11</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">PCR
+
-
reaction was carried out using the primer.</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>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">dNTP</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
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-
<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>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Primer
+
-
mix</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">1ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
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-
<TD WIDTH=147>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Genome
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-
DNA</FONT></SPAN></FONT></P>
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-
</TD>
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-
<TD WIDTH=147>
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-
<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>
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-
<TD WIDTH=147>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">KOD-FX</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
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-
<TD WIDTH=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
+
-
</TD>
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-
<TD WIDTH=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">26.5ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
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-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Electrophoresed
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-
in 1% agarose gel. Verified as </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1.</I></FONT></SPAN></FONT></P>
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Purified
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-
PCR products of </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1.</I></FONT></SPAN></FONT></P>
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<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>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
and pET-15b with Xho1 and Bpu11021 at 37℃ for 2 hours.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<TABLE WIDTH=167 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=96>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT></SPAN></FONT></P>
+
-
</TD>
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-
<TD WIDTH=42>
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-
<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=96>
+
-
<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=96>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu11021</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=96>
+
-
<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=96>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2NEB
+
-
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>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Added
+
-
1ul of BAP to the solution of pET-15b and incubated it at 37℃ for 30
+
-
minutes.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligated
+
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of pET-15b with </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1
+
-
</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">and transformed.</FONT></SPAN></FONT></P>
+
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Applied
+
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pET-15b and </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
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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
+
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and purified them.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
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</P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
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</P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
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12th</B></FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
+
-
32 colonies of </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1
+
-
</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
+
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13th</B></FONT></SPAN></FONT></P>
+
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<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 at 37℃ for 5 hours.</FONT></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></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Digested
+
-
plasmid DNA with Xho1 and Bpu11021 at 37℃ for 2 hours.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=206 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1
+
-
</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">into pET-15b</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=135>
+
-
<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=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu11021</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=135>
+
-
<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=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2NEB
+
-
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>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Applied
+
-
pET-15b to the 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">No
+
-
band was detected.</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 digested with Xho1 and Bpu11021 at 37℃ (overnight).</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<TABLE WIDTH=206 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1
+
-
</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">into pET-15b</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=135>
+
-
<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=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu11021</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=135>
+
-
<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=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2NEB
+
-
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>
+
-
</TABLE>
+
-
<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>September
+
-
14</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>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
and pET-15b were Applied</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>
+
-
</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">to electrophoresis to
+
-
the agarose gel.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">No
+
-
band was detected.</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>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
and pET-15b with Bpu11021 at 37℃ overnight.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<TABLE WIDTH=153 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">26ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu11021</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<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=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu
+
-
Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<TABLE WIDTH=153 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">26ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu11021</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<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=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu
+
-
Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</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>September
+
-
15</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
+
-
</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
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">Digested
+
-
</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
and pET-15b with Xho1.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=153 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">26ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho1</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<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=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu
+
-
Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<TABLE WIDTH=153 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">26ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Xho11021</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<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=83>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu
+
-
Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=40>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">3ul</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">pET-15b
+
-
and </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
were applied to electrophoresis to the Blue gel and purified.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligated
+
-
</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
with pET-15b and transformed them 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
+
-
16th</B></FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
+
-
32 colonies of </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1
+
-
</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
+
-
18th</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 at 37℃
+
-
(overnight).</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
+
-
19</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 purified and digested with Xho1 and Bpu11021 at 37℃ for 2
+
-
hours.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=206 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1
+
-
</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">into pET-15b</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=135>
+
-
<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=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu11021</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=135>
+
-
<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=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2NEB
+
-
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>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
and pET-15b were applied</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>
+
-
</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">to electrophoresis to
+
-
the agarose gel.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">No
+
-
band was detected.</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>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
and pET-15b with Xho1 and Blp1.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=206 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=135>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></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=135>
+
-
<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=135>
+
-
<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>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=135>
+
-
<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=135>
+
-
<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>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<TABLE WIDTH=206 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=135>
+
-
<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">15.5ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=135>
+
-
<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=135>
+
-
<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>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=135>
+
-
<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=135>
+
-
<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>
+
-
</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">Added
+
-
1ul of BAP to the solution of pET-15b and incubated it at 37℃ for 30
+
-
minutes.</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 them.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">The
+
-
</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1 </I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">was
+
-
ligated with pET-15b, and transformed </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>into</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
</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
+
-
20</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
+
-
64 colonies of </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1
+
-
</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"><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
+
-
21st</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 colonies and cultured in the LB in 3ml LB medium with ampicillin
+
-
at 37℃ (overnight).</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>September
+
-
22nd</B></FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Miniprepped
+
-
plasmid DNA and digested it with Xho1 and Blp1 at 37℃ for 2 hours.</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>
+
-
ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt"> into pET-15b to
+
-
the 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">No
+
-
band was detected.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 3<sup>rd</sup></span></b></p>
-
</div>
+
<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'>No appropriate
 +
plasmid DNA was detected.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested <i>ATF1</i>
 +
and pET-15b with <i>Xho</i>I and <i>Bpu</i>1102I at 37</span><span
 +
style='font-size:11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C overnight.</span></p>
 +
 
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none'>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;border:solid windowtext 1.0pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>ATF1</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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 2</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;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=111 valign=top style='width:83.2pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='height:20.15pt'>
 +
  <td width=111 valign=top style='width:83.2pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt;height:20.15pt'>
 +
  <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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;height:20.15pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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 2</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 4<sup>th&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </sup></span></b></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'>No pET-15b was
 +
detected.</span></p>
 +
 
 +
<p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i><span
 +
lang=EN-US style='font-size:11.0pt'> was extracted from the blue gel.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 11<sup>th</sup></span></b></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>PCR reaction was
 +
carried out using the primer. </span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>(Forward : CGATCCTCGAGTGGCCTGGTATTGTCATCGCGTTG</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Reverse : CGATCGCTCAGCAACCAACCAAAGCCGAGGGAGTG)</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&micro;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&micro;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&micro;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&micro;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&micro;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Electrophoresed in
 +
1% agarose gel. Verified as <i>ATF1.</i></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified the PCR
 +
products of <i>ATF1.</i></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested <i>ATF1</i>
 +
and pET-15b with <i>Xho</i>I and <i>Bpu</i>1102I at 37</span><span
 +
style='font-size:11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C for 2 hours.</span></p>
 +
 
 +
<p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></i></p>
 +
 
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none'>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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'>ATF1</span></i></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=111 valign=top style='width:83.2pt;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 2</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Added 1&micro;L of BAP
 +
to the solution of pET-15b and incubated it at 37</span><span style='font-size:
 +
11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US style='font-size:
 +
11.0pt'>C for 30 minutes.</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 them.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligated pET-15b and
 +
<i>ATF1 </i>and transformed.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 12th</span></b></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked 32 colonies
 +
of <i>ATF1 </i>transformants.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 13th</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 3mL LB medium with ampicillin at 37</span><span
 +
style='font-size:11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C for 5 hours.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Miniprepped plasmid
 +
DNA.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested plasmid
 +
DNA with <i>Xho</i>I and <i>Bpu</i>1102I at 37</span><span style='font-size:
 +
11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US style='font-size:
 +
11.0pt'>C for 2 hours.</span></p>
 +
 
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none'>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>ATF1 </span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>into pET-15b</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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 2</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied pET-15b to
 +
the agarose gel electrophoresis.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>No band was
 +
detected.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Plasmid DNA was
 +
digested with <i>Xho</i>I and <i>Bpu</i>1102I at 37</span><span
 +
style='font-size:11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C (overnight).</span></p>
 +
 
 +
<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=150 valign=top style='width:112.25pt;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'>ATF1 </span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>into pET-15b</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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 2</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;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>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 14<sup>th</sup></span></b></p>
 +
 
 +
<p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i><span
 +
lang=EN-US style='font-size:11.0pt'> and pET-15b were applied<i> </i>to
 +
electrophoresis to the agarose gel.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>No band was
 +
detected.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested <i>ATF1</i>
 +
and pET-15b with <i>Bpu</i>1102I at 37</span><span style='font-size:11.0pt;
 +
font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US style='font-size:11.0pt'>C
 +
overnight.</span></p>
 +
 
 +
<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=98 valign=top style='width:73.65pt;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'>ATF1</span></i></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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'>26&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=98 valign=top style='width:73.65pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=98 valign=top style='width:73.65pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I Buffer</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;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=98 valign=top style='width:73.65pt;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=55 valign=top style='width:41.2pt;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'>26&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=98 valign=top style='width:73.65pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=98 valign=top style='width:73.65pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I Buffer</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;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>&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'>Purified <i>ATF1</i>
 +
and pET-15b.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested <i>ATF1</i>
 +
and pET-15b with <i>Xho</i>I.</span></p>
 +
 
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none'>
 +
<tr>
 +
  <td width=98 valign=top style='width:73.65pt;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'>ATF1</span></i></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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'>26&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=98 valign=top style='width:73.65pt;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'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=98 valign=top style='width:73.65pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I Buffer</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;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=98 valign=top style='width:73.65pt;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=55 valign=top style='width:41.2pt;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'>26&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=98 valign=top style='width:73.65pt;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'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=98 valign=top style='width:73.65pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I Buffer</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;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'>pET-15b and <i>ATF1</i>
 +
were applied to electrophoresis in the Blue gel and purified.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligated <i>ATF1</i>
 +
and pET-15b and transformed them into <i>E. coli </i>cells.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 16th</span></b></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked 32 colonies
 +
of <i>ATF1 </i>transformants.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 18th</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 at 37</span><span
 +
style='font-size:11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C (overnight).</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&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><span lang=EN-US style='font-size:11.0pt'>plasmid DNA was
 +
purified and digested with <i>Xho</i>I and<i> Bpu</i>1102I at 37</span><span
 +
style='font-size:11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C for 2 hours.</span></p>
 +
 
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none'>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>ATF1 </span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>into pET-15b</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Bpu</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>1102I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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 2</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 
 +
<p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i><span
 +
lang=EN-US style='font-size:11.0pt'> and pET-15b were applied<i> </i>to
 +
electrophoresis to the agarose gel.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>No band was
 +
detected.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested <i>ATF1</i>
 +
and pET-15b with <i>Xho</i>I and <i>Blp</i>I.</span></p>
 +
 
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none'>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>ATF1</span></i></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Blp</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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=55 valign=top style='width:41.2pt;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&micro;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=150 valign=top style='width:112.25pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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'>Blp</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I</span></p>
 +
  </td>
 +
  <td width=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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=55 valign=top style='width:41.2pt;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&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=150 valign=top style='width:112.25pt;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=55 valign=top style='width:41.2pt;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&micro;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'>Added 1&micro;L of BAP
 +
to the solution of pET-15b and incubated it at 37</span><span style='font-size:
 +
11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US style='font-size:
 +
11.0pt'>C for 30 minutes.</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 them.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>The <i>ATF1 </i>was
 +
ligated with pET-15b, and transformed <i>into</i> <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='color:red'>&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 <i>ATF1 </i>transformants.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&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 colonies
 +
and cultured in the LB in 3mL LB medium with ampicillin at 37</span><span
 +
style='font-size:11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C (overnight).</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>September 22nd</span></b></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Miniprepped plasmid
 +
DNA and digested it with <i>Xho</i>I and<i> Blp</i>I at 37</span><span
 +
style='font-size:11.0pt;font-family:"MS 明朝","serif"'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C for 2 hours.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied<i> ATF1</i>
 +
into pET-15b to the agarose gel electrophoresis.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>No band was
 +
detected.</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>
</div>
</div>
-
</html>
+
</div>

Latest revision as of 09:19, 27 September 2013



ATF1

September 1st

Picked 32 colonies of ATF1 transformants.

 

September 3rd

Checked the colonies by colony cracking.

No appropriate plasmid DNA was detected.

Digested ATF1 and pET-15b with XhoI and Bpu1102I at 37˚C overnight.

ATF1

24µL

XhoI

1µL

Bpu1102I

1µL

BSA

1µL

NEB Buffer 2

3µL

 

pET-15b

33µL

XhoI

1µL

Bpu1102I

1µL

BSA

1µL

NEB Buffer 2

4µL

 

 

September 4th       

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

No pET-15b was detected.

ATF1 was extracted from the blue gel.

 

 

September 11th

PCR reaction was carried out using the primer.

(Forward : CGATCCTCGAGTGGCCTGGTATTGTCATCGCGTTG

Reverse : CGATCGCTCAGCAACCAACCAAAGCCGAGGGAGTG)

Buffer

50µL

dNTP

20µL

Primer mix

1µL

Genome DNA

0.5µL

KOD-FX

2µL

H2O

26.5µL

Electrophoresed in 1% agarose gel. Verified as ATF1.

Purified the PCR products of ATF1.

Digested ATF1 and pET-15b with XhoI and Bpu1102I at 37˚C for 2 hours.

 

ATF1

15.5µL

XhoI

1µL

Bpu1102I

1µL

BSA

0.5µL

NEB Buffer 2

2µL

Added 1µL of BAP to the solution of pET-15b and incubated it at 37˚C for 30 minutes.

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

Isolated and purified them.

Ligated pET-15b and ATF1 and transformed.

 

 

September 12th

Picked 32 colonies of ATF1 transformants.

 

 

September 13th

Checked the colonies by colony cracking.

Picked up the appropriate colonies and cultured in 3mL LB medium with ampicillin at 37˚C for 5 hours.

Miniprepped plasmid DNA.

Digested plasmid DNA with XhoI and Bpu1102I at 37˚C for 2 hours.

ATF1 into pET-15b

15.5µL

XhoI

1µL

Bpu1102I

1µL

BSA

0.5µL

NEB Buffer 2

2µL

Applied pET-15b to the agarose gel electrophoresis.

No band was detected.

Plasmid DNA was digested with XhoI and Bpu1102I at 37˚C (overnight).

 

ATF1 into pET-15b

15.5µL

XhoI

1µL

Bpu1102I

1µL

BSA

0.5µL

NEB Buffer 2

2µL

 

 

 

September 14th

ATF1 and pET-15b were applied to electrophoresis to the agarose gel.

No band was detected.

Digested ATF1 and pET-15b with Bpu1102I at 37˚C overnight.

 

ATF1

26µL

Bpu1102I

1µL

Bpu1102I Buffer

3µL

 

pET-15b

26µL

Bpu1102I

1µL

Bpu1102I Buffer

3µL

 

 

September 15th

Purified ATF1 and pET-15b.

Digested ATF1 and pET-15b with XhoI.

ATF1

26µL

XhoI

1µL

Bpu1102I Buffer

3µL

 

pET-15b

26µL

XhoI

1µL

Bpu1102I Buffer

3µL

 

pET-15b and ATF1 were applied to electrophoresis in the Blue gel and purified.

Ligated ATF1 and pET-15b and transformed them into E. coli cells.

 

 

September 16th

Picked 32 colonies of ATF1 transformants.

 

 

September 18th

Checked the colonies by colony cracking.

Picked up the colonies and cultured in 3mL LB medium with ampicillin at 37˚C (overnight).

 

 

September 19th

plasmid DNA was purified and digested with XhoI and Bpu1102I at 37˚C for 2 hours.

ATF1 into pET-15b

15.5µL

XhoI

1µL

Bpu1102I

1µL

BSA

0.5µL

NEB Buffer 2

2µL

ATF1 and pET-15b were applied to electrophoresis to the agarose gel.

No band was detected.

Digested ATF1 and pET-15b with XhoI and BlpI.

ATF1

15.5µL

XhoI

1µL

BlpI

1µL

BSA

0.5µL

Buffer

2µL

 

pET-15b

15.5µL

XhoI

1µL

BlpI

1µL

BSA

0.5µL

Buffer

2µL

 

Added 1µL of BAP to the solution of pET-15b and incubated it at 37˚C for 30 minutes.

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

Isolated and purified them.

The ATF1 was ligated with pET-15b, and transformed into E. coli cells.

 

 

September 20th

Picked 64 colonies of ATF1 transformants.

 

 

 

 

September 21st

Checked the colonies by colony cracking.

Picked up colonies and cultured in the LB in 3mL LB medium with ampicillin at 37˚C (overnight).

 

September 22nd

Miniprepped plasmid DNA and digested it with XhoI and BlpI at 37˚C for 2 hours.

Applied ATF1 into pET-15b to the agarose gel electrophoresis.

No band was detected.