Team:KIT-Kyoto/Notebook/ATF1/may

From 2013.igem.org

(Difference between revisions)
 
(4 intermediate revisions not shown)
Line 16: Line 16:
<h2>ATF1</h2>
<h2>ATF1</h2>
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT COLOR="#444444"><FONT SIZE=4><B>May
+
<div class=WordSection1 style='layout-grid:18.0pt'>
-
16</B></FONT></FONT><FONT COLOR="#444444"><SUP><FONT SIZE=4><B>th</B></FONT></SUP></FONT><FONT COLOR="#444444"><FONT SIZE=4><B>
+
-
</B></FONT></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 </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1
+
-
</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">gene.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Primers:</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Forward:5&rsquo;-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3&rsquo;</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Reverse:5&rsquo;-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3&rsquo;</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Reaction
+
-
composition is as follows:</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=206 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">dNTP</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Primer
+
-
mix</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">DNA
+
-
sample</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">KOD-FX</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">26.5uL</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">total</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">100uL</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
+
-
could not get any PCR product.</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>May
+
-
17</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP><FONT SIZE=4><B>
+
-
-20</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP><FONT SIZE=4><B>
+
-
</B></FONT></SPAN></FONT>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Changed
+
-
PCR conditions with another DNA template. We performed PCR of the
+
-
</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
gene again.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Primers:</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Forward
+
-
:5&rsquo;-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3&rsquo;</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Reverse:5&rsquo;-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3&rsquo;</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Reaction
+
-
composition is as follows:</FONT></SPAN></FONT></P>
+
-
<TABLE WIDTH=206 BORDER=1 BORDERCOLOR="#00000a" CELLPADDING=7 CELLSPACING=0>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Buffer</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">dNTP</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">Primer
+
-
mix</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">DNA
+
-
sample</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">KOD-FX</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<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=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">H2O</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">26.5uL</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR VALIGN=TOP>
+
-
<TD WIDTH=106>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">total</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=70>
+
-
<P><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">100uL</FONT></SPAN></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
</TABLE>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
+
-
could not get any PCR product.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=4><B>May
+
-
30</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP><FONT SIZE=4><B>
+
-
-31</B></FONT><SUP><FONT SIZE=4><B>th</B></FONT></SUP></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
+
-
transformed a plasmid carrying the </FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I>ATF1</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">
+
-
gene into</FONT><FONT SIZE=2 STYLE="font-size: 11pt"><I> E.coli</I></FONT><FONT SIZE=2 STYLE="font-size: 11pt">.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">This
+
-
plasmid was obtained from iGEM DNA distributions kit 2012 (plate 2,
+
-
o-7).</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT SIZE=2 STYLE="font-size: 11pt">We
+
-
could see approximately 50 colonies and cultured furthermore.</FONT></SPAN></FONT></P>
+
-
<P STYLE="margin-bottom: 0cm"><BR>
+
-
</P>
+
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt;color:#444444;
 +
background:white'>May 16<sup>th</sup> </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>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Primers:</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Forward:5’-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3’</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Reaction
 +
composition is as follows:</span></p>
 +
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='margin-left:42.0pt;border-collapse:collapse;border:none'>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>Buffer</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;border:solid windowtext 1.0pt;
 +
  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>50&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>dNTP</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>20&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>Primer mix</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>DNA sample</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>0.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>KOD-FX</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>2&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>H<sub>2</sub>O</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>26.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>total</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>100&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We could not get any
 +
PCR product.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>May 17<sup>th</sup>
 +
-20<sup>th</sup> </span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Changed PCR conditions
 +
with another DNA template. We performed PCR of the <i>ATF1</i> gene again.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Primers:</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Forward :5’-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3’</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Reaction
 +
composition is as follows:</span></p>
 +
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='margin-left:42.0pt;border-collapse:collapse;border:none'>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>Buffer</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;border:solid windowtext 1.0pt;
 +
  border-left:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>50&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>dNTP</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>20&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>Primer mix</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>DNA sample</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>0.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>KOD-FX</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>2&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>H<sub>2</sub>O</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>26.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr>
 +
  <td width=121 valign=top style='width:91.0pt;border:solid windowtext 1.0pt;
 +
  border-top:none;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>total</span></p>
 +
  </td>
 +
  <td width=85 valign=top style='width:63.8pt;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=MsoListParagraph style='margin-left:0mm'><span lang=EN-US
 +
  style='font-size:11.0pt'>100&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We could not get any
 +
PCR product.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>May 30<sup>th</sup>
 +
-31<sup>th</sup></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We transformed a plasmid
 +
carrying the <i>ATF1</i> gene into<i> E.coli</i>.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>This plasmid was obtained
 +
from iGEM DNA distributions kit 2012 (plate 2, o-7).</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We could see approximately
 +
50 colonies and cultured furthermore.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
</div>
</div>
 +
 +
</div>
</div>

Latest revision as of 08:43, 27 September 2013



KIT-Kyoto


ATF1

May 16th

We performed PCR to amplify the ATF1 gene.

Primers:

Forward:5’-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3’

Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’

Reaction composition is as follows:

Buffer

50µL

dNTP

20µL

Primer mix

1µL

DNA sample

0.5µL

KOD-FX

2µL

H2O

26.5µL

total

100µL

We could not get any PCR product.

 

 

May 17th -20th

Changed PCR conditions with another DNA template. We performed PCR of the ATF1 gene again.

Primers:

Forward :5’-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3’

Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’

Reaction composition is as follows:

Buffer

50µL

dNTP

20µL

Primer mix

1µL

DNA sample

0.5µL

KOD-FX

2µL

H2O

26.5µL

total

100µL

We could not get any PCR product.

 

 

 

May 30th -31th

We transformed a plasmid carrying the ATF1 gene into E.coli.

This plasmid was obtained from iGEM DNA distributions kit 2012 (plate 2, o-7).

We could see approximately 50 colonies and cultured furthermore.

 


Retrieved from "http://2013.igem.org/Team:KIT-Kyoto/Notebook/ATF1/may"