Team:KIT-Kyoto/Notebook/ATF1/august

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<h2>ATF1</h2>
<h2>ATF1</h2>
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 6th</span></b></p>
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
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<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>The <i>ATF1 </i>gene
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6th</B></FONT></SPAN></FONT></P>
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was purified (prepared on 7/19)</span></p>
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">The
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</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1 </I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">gene
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was purified (prepared on 7/19)</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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pET-15b with XhoⅠat 37℃ for 90 minutes.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
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<TR VALIGN=TOP>
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<p class=MsoNormal align=left style='text-align:left;text-autospace:none'><span
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<TD WIDTH=147>
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lang=EN-US style='font-size:11.0pt'>Digested pET-15b with <i>Xho</i>I at 37</span><span
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
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lang=EN-US style='font-size:11.0pt;font-family:メイリオ'> </span><span
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</TD>
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style='font-size:11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span
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<TD WIDTH=147>
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lang=EN-US style='font-size:11.0pt'> for 90 minutes.</span></p>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">88ul</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=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">XhoⅠ
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Buffer</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">10ul</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=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">XhoⅠ</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">2ul</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=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Total</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">100ul</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"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Purified
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pET-15b DNA was dissolved in 34ul of H2O. </FONT></SPAN></FONT>
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</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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pET-15b with Bpu1102Ⅰ at 37℃ for 90 minutes.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
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-
<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">pET-15b</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">34ul</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=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu1102Ⅰ
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Buffer</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">4ul</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=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu1102Ⅰ</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">2ul</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=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Total</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">40ul</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"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Added
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1uL of BAP to the solution and incubated it at 37℃ for 30 minutes.</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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</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 to the blue gel electrophoresis.</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">Results:
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No band 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">Cultivated
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pET-15b in ampicillin (+) liquid LB overnight.</FONT></SPAN></FONT></P>
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<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
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</P>
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<P STYLE="margin-bottom: 0cm; page-break-before: always"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
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7th</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">Minipreped
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pET-15b DNA.</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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pET-15b with XhoⅠ at 37℃ for 3 hours.</FONT></SPAN></FONT></P>
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-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<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">pET-15b</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">88ul</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">XhoⅠ
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Buffer</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">10ul</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">XhoⅠ</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">2ul</FONT></SPAN></FONT></P>
+
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</TD>
+
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</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">Total</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">100ul</FONT></SPAN></FONT></P>
+
-
</TD>
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-
</TR>
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-
</TABLE>
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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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it and dissolved in 26ul of H2O.</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
+
-
pET-15b with Bpu1102Ⅰ at 37℃ for 3 hours.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
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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=147>
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<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=147>
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-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu1102Ⅰ
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Buffer</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">3ul</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">Bpu1102Ⅰ</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>
+
-
<TD WIDTH=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Total</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">30ul</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">Added
+
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1uL of BAP to this solution and incubated it at 37℃ for 30 minutes.</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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</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 to the blue gel electrophoresis.</FONT></SPAN></FONT></P>
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<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
+
-
</P>
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<P STYLE="margin-bottom: 0cm; page-break-before: always"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
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8th</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">Digested
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pET-15b with XhoⅠ at 37℃ overnight.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<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">pET-15b</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">89ul</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">XhoⅠ
+
-
Buffer</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">10ul</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">XhoⅠ</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>
+
-
<TD WIDTH=147>
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Total</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">100ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
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<P STYLE="margin-bottom: 0cm"><BR>
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</P>
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<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
+
-
</P>
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-
<P STYLE="margin-bottom: 0cm; page-break-before: always"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
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9th</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">Purified
+
-
pET-15b (prepared on 8/8) and added 26ul of H2O.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Digested
+
-
pET-15b with Bpu1102Ⅰ at 37℃ for 3 hours.</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">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu1102Ⅰ
+
-
Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu1102Ⅰ</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">Total</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">30ul</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 this solution and incubate 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 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>
+
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<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligation
+
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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">(prepared 7/19,8/6)
+
-
at room temperature for 15 minutes.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
+
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<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">ATF1</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligation
+
-
MIX</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Total</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>
+
-
</TABLE>
+
-
<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">into
+
-
pET-15b was transformed 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=2 STYLE="font-size: 11pt">Cultivated
+
-
transformants on LB ampicillin (+) plate at37℃ overnight.</FONT></SPAN></FONT></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>August
+
-
10th</B></FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Picked
+
-
48 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>August
+
-
12th</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 at 37℃ for 4 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"><BR>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
+
-
13th</B></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 (prepared on 8/12) with HindⅢ at 37℃ for 90 minutes.</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">Plasmid
+
-
DNA</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=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">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">12ul</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">HindⅢ</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">Total</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>
+
-
</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">Digested
+
-
plasmid DNA (prepared on 8/12) with HindⅢ at 37℃ for 90 minutes.</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">Plasmid
+
-
DNA</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=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">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">7.25ul</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">HindⅢ</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">0.75ul</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">Total</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>
+
-
</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">Again,
+
-
picked up the appropriate colonies and cultured in the 3ml LB medium
+
-
with ampicillin at 37℃.</FONT></SPAN></FONT></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"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
+
-
26th</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 the ATF1 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"><BR>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
+
-
27th</B></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 (prepared on 8/26) and added 30ul of H2O.</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">
+
-
with HindⅢ.</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">ATF1l</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=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">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">HindⅢ</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">11ul</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">Digested
+
-
pET-15b and </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
with XhoⅠ.</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">ATF1</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">25ul</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">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">XhoⅠ</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>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</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">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">100ul</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">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">11.3ul</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">XhoⅠ</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>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Purified
+
-
it and added 25ul of H2O.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Digested
+
-
pET-15b and </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1 </I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">with
+
-
Bpu1102Ⅰ.</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">ATF1</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">25ul</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">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu1102Ⅰ</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>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</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">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">25ul</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">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Bpu1102Ⅰ</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>
+
-
</TABLE>
+
-
<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">Purified
+
-
PCR products.</FONT></SPAN></FONT></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"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
+
-
28th</B></FONT></SPAN></FONT></P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><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">(prepared
+
-
on 8/27) with EcoRⅠ .</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">ATF1l</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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 ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">EcoRⅠ</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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 ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Applied
+
-
it to the agarose gel electrophoresis.</FONT></SPAN></FONT></P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><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">(prepared
+
-
on 8/27) and pET-15b with XhoⅠ.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">ATF1</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">25ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">XhoⅠ</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
+
-
</P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">100ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">11.3ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">XhoⅠ</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">2ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
+
-
</P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
+
-
</P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
+
-
29th</B></FONT></SPAN></FONT></P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><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">and
+
-
pET-15b to the blue gel electrophoresis.</FONT></SPAN></FONT></P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligated
+
-
it.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">ATF1</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligation
+
-
MIX</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">10ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Carried
+
-
out transformation.</FONT></SPAN></FONT></P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
+
-
</P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><BR>
+
-
</P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>August
+
-
30th</B></FONT></SPAN></FONT></P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Checked
+
-
the colonies by colony cracking.</FONT></SPAN></FONT></P>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
+
-
performed PCR to amplify the ATF1 gene.</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=324 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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 ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">dNTP</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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 ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Primer
+
-
mix</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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 ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Genome
+
-
DNA</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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 ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">KOD-FX</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><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 ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<P ALIGN=LEFT STYLE="widows: 2; orphans: 2"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">26.5ul</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
-
<P ALIGN=LEFT STYLE="margin-bottom: 0cm; widows: 2; orphans: 2"><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>August
+
-
31</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">Purified
+
-
PCR products and dissolved 30ul 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 and added 33ul of H2O.</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 XhoⅠ and Bpu1102Ⅰ.</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">NEB
+
-
Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">ATF1</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</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">XhoⅠ</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">Bpu1102Ⅰ</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>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</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">NEB2
+
-
Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">BSA</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">XhoⅠ</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">Bpu1102Ⅰ</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>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Added
+
-
1uL of BAP to pET-15b and incubated them 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 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>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
(prep).</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">ATF1</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">pET-15b</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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=147>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Ligation
+
-
MIX</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=147>
+
-
<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>ATF1</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"><BR>
+
-
</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'>pET-15b</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'>88</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I Buffer</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'>10</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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'>Total</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'>100</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified pET-15b DNA
 +
was dissolved in 34</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 align=left style='text-align:left;text-autospace:none'><span
 +
lang=EN-US style='font-size:11.0pt'>Digested pET-15b with <i>Bpu</i>1102I at 37</span><span
 +
lang=EN-US style='font-size:11.0pt;font-family:メイリオ'> </span><span
 +
style='font-size:11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span
 +
lang=EN-US style='font-size:11.0pt'> for 90 minutes.</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'>pET-15b</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'>34</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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'>4</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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'>Total</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'>40</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal align=left style='text-align:left;text-autospace:none'><span
 +
lang=EN-US style='font-size:11.0pt'>Added 1uL of BAP to the solution and
 +
incubated it at 37</span><span lang=EN-US style='font-size:11.0pt;font-family:
 +
メイリオ'> </span><span style='font-size:11.0pt;font-family:メイリオ'>&#730;<span
 +
lang=EN-US>C </span></span><span lang=EN-US style='font-size:11.0pt'>for 30
 +
minutes.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied <i>ATF1</i>
 +
and pET-15b to the blue gel electrophoresis.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Results: No band
 +
was detected.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Cultivated pET-15b
 +
in 3mL LB medium with ampicillin overnight.</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'>August 7th</span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Minipreped pET-15b
 +
DNA.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b
 +
with <i>Xho</i>I at 37</span><span style='font-size:11.0pt;font-family:メイリオ'>&#730;<span
 +
lang=EN-US>C</span></span><span lang=EN-US style='font-size:11.0pt'> for 3
 +
hours.</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'>pET-15b</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'>88</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I Buffer</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'>10</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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'>Total</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'>100</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified it and dissolved
 +
in 26</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><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b
 +
with <i>Bpu</i>1102I at 37</span><span style='font-size:11.0pt;font-family:
 +
メイリオ'>&#730;<span lang=EN-US>C</span></span><span lang=EN-US style='font-size:11.0pt'>
 +
for 3 hours.</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'>pET-15b</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'>26</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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'>3</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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'>Total</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'>30</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Added 1</span><span
 +
lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
11.0pt'>L of BAP to this solution and incubated it at 37</span><span
 +
style='font-size:11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span
 +
lang=EN-US style='font-size:11.0pt'> for 30 minutes.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied <i>ATF1</i>
 +
and pET-15b to the blue gel electrophoresis.</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'>August 8th</span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b
 +
with <i>Xho</i>I at 37</span><span style='font-size:11.0pt;font-family:メイリオ'>&#730;<span
 +
lang=EN-US>C</span></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=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'>pET-15b</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'>89</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>I Buffer</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'>10</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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'>Total</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'>100</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<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'>August 9th</span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified pET-15b (prepared
 +
on 8/8) and added 26ul of H<sub>2</sub>O.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b
 +
with <i>Bpu</i>1102I at 37</span><span style='font-size:11.0pt;font-family:
 +
メイリオ'>&#730;<span lang=EN-US>C</span></span><span lang=EN-US style='font-size:11.0pt'>
 +
for 3 hours.</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'>pET-15b</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'>26</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><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=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'>3</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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'>Total</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'>30</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Added 1</span><span
 +
lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
11.0pt'>L of BAP to this solution and incubate it at 37</span><span
 +
style='font-size:11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span
 +
lang=EN-US style='font-size:11.0pt'> for 30 minutes.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied pET-15b 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 pET-15b
 +
and <i>ATF1 </i>(prepared 7/19,8/6) at room temperature for 15 minutes.</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><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></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'>5</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>pET-15b</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'>5</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Ligation 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'>10</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Total</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:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>The <i>ATF1 </i>into
 +
pET-15b was transformed into <i>E.coli </i>cells.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Cultivated
 +
transformants on LB plate with ampicillin at37</span><span style='font-size:
 +
11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span lang=EN-US
 +
style='font-size:11.0pt'> overnight.</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'>August 10th</span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked 48 colonies
 +
of <i>ATF1 </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'>August 12th</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 at 37</span><span
 +
style='font-size:11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span
 +
lang=EN-US style='font-size:11.0pt'> for 4 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'>&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'>August 13th</span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested plasmid
 +
DNA (prepared on 8/12) with<i> Hin</i>dIII at 37</span><span style='font-size:
 +
11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span lang=EN-US
 +
style='font-size:11.0pt'> for 90 minutes.</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'>Plasmid DNA</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'>5</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Buffer</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:
 +
  11.0pt'>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'>12</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><i><span lang=EN-US style='font-size:11.0pt'>Hin</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>dIII</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:
 +
  11.0pt'>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'>Total</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:
 +
  11.0pt'>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'>Digested plasmid
 +
DNA (prepared on 8/12) with <i>Hin</i>dIII at 37</span><span style='font-size:
 +
11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span lang=EN-US
 +
style='font-size:11.0pt'> for 90 minutes.</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'>Plasmid DNA</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'>10</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Buffer</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:
 +
  11.0pt'>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'>7.25</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><i><span lang=EN-US style='font-size:11.0pt'>Hin</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>dIII</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'>0.75</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Total</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:
 +
  11.0pt'>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'>Again, picked up the
 +
appropriate colonies and cultured in the 3mL LB medium with ampicillin at 37</span><span
 +
style='font-size:11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span
 +
lang=EN-US style='font-size:11.0pt'>.</span></p>
 +
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>&nbsp;</span></b></p>
 +
 +
<b><span lang=EN-US style='font-size:14.0pt;font-family:"Century","serif"'><br
 +
clear=all style='page-break-before:always'>
 +
</span></b>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US
 +
style='font-size:14.0pt'>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 26th</span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR
 +
to amplify the <i>ATF1 </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:
 +
  11.0pt'>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:
 +
  11.0pt'>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:
 +
  11.0pt'>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:
 +
  11.0pt'>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:
 +
  11.0pt'>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:
 +
  11.0pt'>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'>August 27th</span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified PCR
 +
products (prepared on 8/26) and added 30</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><span lang=EN-US style='font-size:11.0pt'>Digested <i>ATF1</i>
 +
with <i>Hin</i>dIII.</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><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></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'>5</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Buffer</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:
 +
  11.0pt'>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><i><span lang=EN-US style='font-size:11.0pt'>Hin</span></i><span
 +
  lang=EN-US style='font-size:11.0pt'>dIII</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:
 +
  11.0pt'>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'>11</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b
 +
and <i>ATF1</i> 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=162 valign=top style='width:121.45pt;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'>ATF1</span></i></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'>25</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Buffer</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'>3</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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=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'>pET-15b</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'>100</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Buffer</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'>11.3</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified it and
 +
added 25</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><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b
 +
and <i>ATF1 </i>with <i>Bpu</i>1102I.</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><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></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'>25</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Buffer</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'>3</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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=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'>pET-15b</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'>25</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Buffer</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'>3</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<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'>Purified PCR
 +
products.</span></p>
 +
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US
 +
style='font-size:14.0pt'>August 28th</span></b></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>Digested <i>ATF1 </i>(prepared on 8/27) with <i>Eco</i>RI.</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 align=left style='text-align:left'><i><span lang=EN-US
 +
  style='font-size:11.0pt'>ATF1</span></i></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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><span lang=EN-US
 +
  style='font-size:11.0pt'>Buffer</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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><i><span lang=EN-US
 +
  style='font-size:11.0pt'>Eco</span></i><span lang=EN-US style='font-size:
 +
  11.0pt'>RI</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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><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 align=left style='text-align:left'><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:11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>Applied it to the agarose gel electrophoresis.</span></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>Digested <i>ATF1 </i>(prepared on 8/27) 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=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal align=left style='text-align:left'><i><span lang=EN-US
 +
  style='font-size:11.0pt'>ATF1</span></i></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 align=left style='text-align:left'><span lang=EN-US
 +
  style='font-size:11.0pt'>25</span><span lang=ja style='font-size:11.0pt'>&micro;</span><span
 +
  lang=EN-US style='font-size:11.0pt'>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 align=left style='text-align:left'><span lang=EN-US
 +
  style='font-size:11.0pt'>Buffer</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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><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=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 align=left style='text-align:left'><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:11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><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=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
  style='font-size:11.0pt'>pET-15b</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 align=left style='text-align:left'><span lang=EN-US
 +
  style='font-size:11.0pt'>100</span><span lang=ja style='font-size:11.0pt'>&micro;</span><span
 +
  lang=EN-US style='font-size:11.0pt'>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 align=left style='text-align:left'><span lang=EN-US
 +
  style='font-size:11.0pt'>Buffer</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 align=left style='text-align:left'><span lang=EN-US
 +
  style='font-size:11.0pt'>11.3</span><span lang=ja style='font-size:11.0pt'>&micro;</span><span
 +
  lang=EN-US style='font-size:11.0pt'>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 align=left style='text-align:left'><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=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 align=left style='text-align:left'><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:11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US
 +
style='font-size:14.0pt'>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US
 +
style='font-size:14.0pt'>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US
 +
style='font-size:14.0pt'>August 29th</span></b></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>Applied <i>ATF1 </i>and pET-15b to the blue gel electrophoresis.</span></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>Ligated it.</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 align=left style='text-align:left'><i><span lang=EN-US
 +
  style='font-size:11.0pt'>ATF1</span></i></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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><span lang=EN-US
 +
  style='font-size:11.0pt'>pET-15b</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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><span lang=EN-US
 +
  style='font-size:11.0pt'>Ligation&nbsp; 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 align=left style='text-align:left'><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:11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>Carried out transformation.</span></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US
 +
style='font-size:14.0pt'>August 30th</span></b></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>Checked the colonies by colony cracking.</span></p>
 +
 +
<p class=MsoNormal align=left style='text-align:left'><span lang=EN-US
 +
style='font-size:11.0pt'>We performed PCR to amplify the <i>ATF1</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 align=left style='text-align:left'><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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><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 align=left style='text-align:left'><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:11.0pt'>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 align=left style='text-align:left'><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 align=left style='text-align:left'><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:11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<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><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'>August 31<sup>st</sup></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified PCR
 +
products and dissolved 30</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><span lang=EN-US style='font-size:11.0pt'>Purified pET-15b
 +
and added 33</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><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.</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'>NEB Buffer 2</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'>3</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></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'>24</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>BSA</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:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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=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'>NEB Buffer 2</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'>4</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>pET-15b</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'>33</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>BSA</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:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>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><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=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:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Added 1</span><span
 +
lang=EN-US style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
11.0pt'>L of BAP to pET-15b and incubated them at 37</span><span
 +
style='font-size:11.0pt;font-family:メイリオ'>&#730;<span lang=EN-US>C</span></span><span
 +
lang=EN-US style='font-size:11.0pt'> for 30 minutes.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied pET-15b 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>ATF1</i> (prep).</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><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></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'>10</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>pET-15b</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'>10</span><span
 +
  lang=ja style='font-size:11.0pt'>&micro;</span><span lang=EN-US style='font-size:
 +
  11.0pt'>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'>Ligation&nbsp;
 +
  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;
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  padding:0mm 5.4pt 0mm 5.4pt'>
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  <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:
 +
  11.0pt'>L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Transformed <i>ATF1</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><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
</div>
</div>
</div>
</div>

Latest revision as of 09:18, 27 September 2013



ATF1

August 6th

The ATF1 gene was purified (prepared on 7/19)

Digested pET-15b with XhoI at 37 ˚C for 90 minutes.

pET-15b

88µL

XhoI Buffer

10µL

XhoI

2µL

Total

100µL

Purified pET-15b DNA was dissolved in 34µL of H2O.

Digested pET-15b with Bpu1102I at 37 ˚C for 90 minutes.

pET-15b

34µL

Bpu1102I Buffer

4µL

Bpu1102I

2µL

Total

40µL

Added 1uL of BAP to the solution and incubated it at 37 ˚C for 30 minutes.

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

Results: No band was detected.

Cultivated pET-15b in 3mL LB medium with ampicillin overnight.


 

August 7th

Minipreped pET-15b DNA.

Digested pET-15b with XhoI at 37˚C for 3 hours.

pET-15b

88µL

XhoI Buffer

10µL

XhoI

2µL

Total

100µL

Purified it and dissolved in 26µL of H2O.

Digested pET-15b with Bpu1102I at 37˚C for 3 hours.

pET-15b

26µL

Bpu1102I Buffer

3µL

Bpu1102I

1µL

Total

30µL

Added 1µL of BAP to this solution and incubated it at 37˚C for 30 minutes.

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


 

August 8th

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

pET-15b

89µL

XhoI Buffer

10µL

XhoI

1µL

Total

100µL

 


 

August 9th

Purified pET-15b (prepared on 8/8) and added 26ul of H2O.

Digested pET-15b with Bpu1102I at 37˚C for 3 hours.

pET-15b

26

Bpu1102I Buffer

3µL

Bpu1102I

1µL

Total

30µL

Added 1µL of BAP to this solution and incubate it at 37˚C for 30 minutes.

Applied pET-15b to the blue gel electrophoresis.

Isolated and Purified it.

Ligation pET-15b and ATF1 (prepared 7/19,8/6) at room temperature for 15 minutes.

ATF1

5µL

pET-15b

5µL

Ligation MIX

10µL

Total

20µL

The ATF1 into pET-15b was transformed into E.coli cells.

Cultivated transformants on LB plate with ampicillin at37˚C overnight.


 

August 10th

Picked 48 colonies of ATF1 transformants.

 

 

August 12th

Checked the colonies by colony cracking.

Picked up the appropriate colonies and cultured in the LB in 3mL ampicillin(+) medium at 37˚C for 4 hours.

Miniprepped plasmid DNA.

 

 

August 13th

Digested plasmid DNA (prepared on 8/12) with HindIII at 37˚C for 90 minutes.

Plasmid DNA

5µL

Buffer

2µL

H2O

12µL

HindIII

1µL

Total

20µL

Applied pET-15b to the agarose gel electrophoresis.

Digested plasmid DNA (prepared on 8/12) with HindIII at 37˚C for 90 minutes.

Plasmid DNA

10µL

Buffer

2µL

H2O

7.25µL

HindIII

0.75µL

Total

20µL

Applied pET-15b to the agarose gel electrophoresis.

Again, picked up the appropriate colonies and cultured in the 3mL LB medium with ampicillin at 37˚C.

 

 


 

August 26th

We performed PCR to amplify the ATF1 gene.

Buffer

50µL

dNTP

20µL

Primer mix

1µL

Genome DNA

0.5µL

KOD-FX

2µL

H2O

26.5µL

 

 

August 27th

Purified PCR products (prepared on 8/26) and added 30µL of H2O.

Digested ATF1 with HindIII.

ATF1

5µL

Buffer

2µL

HindIII

2µL

H2O

11µL

Digested pET-15b and ATF1 with XhoI.

ATF1

25µL

Buffer

3µL

XhoI

2µL

 

pET-15b

100µL

Buffer

11.3µL

XhoI

2µL

Purified it and added 25µL of H2O.

Digested pET-15b and ATF1 with Bpu1102I.

ATF1

25µL

Buffer

3µL

Bpu1102I

2µL

 

pET-15b

25µL

Buffer

3µL

Bpu1102I

2µL

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

Purified PCR products.

 

 

August 28th

Digested ATF1 (prepared on 8/27) with EcoRI.

ATF1

5µL

Buffer

1µL

EcoRI

2µL

H2O

2µL

Applied it to the agarose gel electrophoresis.

Digested ATF1 (prepared on 8/27) and pET-15b with XhoI.

ATF1

25µL

Buffer

3µL

XhoI

2µL

 

pET-15b

100µL

Buffer

11.3µL

XhoI

2µL

 

 

August 29th

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

Ligated it.

ATF1

10µL

pET-15b

10µL

Ligation  MIX

10µL

Carried out transformation.

 

 

August 30th

Checked the colonies by colony cracking.

We performed PCR to amplify the ATF1 gene.

Buffer

50µL

dNTP

20µL

Primer mix

1µL

Genome DNA

0.5µL

KOD-FX

2µL

H2O

26.5µL

 

 

 

August 31st

Purified PCR products and dissolved 30µL of H2O.

Purified pET-15b and added 33µL of H2O.

Digested ATF1 and pET-15b with XhoI and Bpu1102I.

NEB Buffer 2

3µL

ATF1

24µL

BSA

1µL

XhoI

1µL

Bpu1102I

1µL

 

NEB Buffer 2

4µL

pET-15b

33µL

BSA

1µL

XhoI

1µL

Bpu1102I

1µL

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

Applied pET-15b to the blue gel electrophoresis.

Isolated and purified it.

Ligation of pET-15b with ATF1 (prep).

ATF1

10µL

pET-15b

10µL

Ligation  MIX

10µL

Transformed ATF1 into pET-15b E. coli cells.