Team:KIT-Kyoto/Notebook/ATF1/june

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<h2>ATF1</h2>
<h2>ATF1</h2>
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 4<sup>th</sup></span></b><b
 +
style='mso-bidi-font-weight:normal'><span lang=EN-US style='font-size:11.0pt'></span></b></p>
 +
 +
<p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>Minipreped</span></span><span
 +
lang=EN-US style='font-size:11.0pt'> <span class=GramE><i style='mso-bidi-font-style:
 +
normal'>ATF1</i>(</span>prepared on 5/31).</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 style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 6<sup>th</sup></span></b></p>
 +
 +
<p class=MsoNormal><span class=GramE><span lang=EN-US style='font-size:11.0pt'>Performed
 +
PCR to amplify the <i>ATF1</i> gene.</span></span><span lang=EN-US
 +
style='font-size:11.0pt'></span></p>
 +
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
 +
mso-yfti-tbllook:1184;mso-padding-alt:0mm 5.4pt 0mm 5.4pt'>
 +
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  mso-border-alt:solid windowtext .5pt;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=51 valign=top style='width:51.4pt;border:solid windowtext 1.0pt;
 +
  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
 +
  solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>50&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:1'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>dNTP</span></span><span
 +
  lang=EN-US style='font-size:11.0pt'></span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:2'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:3'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:4'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Forward:
 +
TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Reverse:
 +
ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>35cycle</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 style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 7th</span></b><b
 +
style='mso-bidi-font-weight:normal'><span lang=EN-US style='font-size:11.0pt'></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Electrophoresed
 +
PCR products in 1% <span class=SpellE>agarose</span> gel. No DNA was detected.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 10<sup>th</sup></span></b></p>
 +
 +
<p class=MsoNormal><span class=GramE><span lang=EN-US style='font-size:11.0pt'>Performed
 +
PCR to amplify the <i>ATF1</i> gene.</span></span><span lang=EN-US
 +
style='font-size:11.0pt'></span></p>
 +
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
 +
mso-yfti-tbllook:1184;mso-padding-alt:0mm 5.4pt 0mm 5.4pt'>
 +
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  mso-border-alt:solid windowtext .5pt;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=51 valign=top style='width:51.4pt;border:solid windowtext 1.0pt;
 +
  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
 +
  solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>50&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:1'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>dNTP</span></span><span
 +
  lang=EN-US style='font-size:11.0pt'></span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:2'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:3'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:4'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes'>
 +
  <td width=90 valign=top style='width:89.9pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>Predenature</span></span><span
 +
lang=EN-US style='font-size:11.0pt'> 94</span><span style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;
 +
mso-fareast-mso-fareast-theme-font:minor-fareast;
 +
mso-hansi-theme-font:minor-latin'>&#730;</span><span
 +
lang=EN-US style='font-size:11.0pt'>C 2min</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Denature 98</span><span
 +
style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-mso-fareast-theme-font:
 +
minor-fareast;mso-hansi-theme-font:minor-latin'>&#730;</span><span
 +
lang=EN-US style='font-size:11.0pt'>C 10sec</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Annealing 51</span><span
 +
style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-mso-fareast-theme-font:
 +
minor-fareast;mso-hansi-theme-font:minor-latin'>&#730;</span><span
 +
lang=EN-US style='font-size:11.0pt'>C 30sec</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Extension 68</span><span
 +
style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-mso-fareast-theme-font:
 +
minor-fareast;mso-hansi-theme-font:minor-latin'>&#730;</span><span
 +
lang=EN-US style='font-size:11.0pt'>C 1min30sec</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>(30 cycles)</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 11<sup>th</sup></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Electrophoresed
 +
PCR products in 1% <span class=SpellE>agarose</span> gel.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We detected the
 +
correct size of DNA band.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR
 +
to amplify the <i style='mso-bidi-font-style:normal'>ATF1</i> gene.</span></p>
 +
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
 +
mso-yfti-tbllook:1184;mso-padding-alt:0mm 5.4pt 0mm 5.4pt'>
 +
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  mso-border-alt:solid windowtext .5pt;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=51 valign=top style='width:51.4pt;border:solid windowtext 1.0pt;
 +
  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
 +
  solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>50&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:1'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>dNTP</span></span><span
 +
  lang=EN-US style='font-size:11.0pt'></span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:2'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:3'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><i style='mso-bidi-font-style:normal'><span lang=EN-US
 +
  style='font-size:11.0pt'>ATF1</span></i><span lang=EN-US style='font-size:
 +
  11.0pt'>(PCR product)</span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:4'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>Predenature</span></span><span
 +
lang=EN-US style='font-size:11.0pt'> 94</span><span style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;
 +
mso-fareast-mso-fareast-theme-font:minor-fareast;
 +
mso-hansi-theme-font:minor-latin'>&#730;</span><span
 +
lang=EN-US style='font-size:11.0pt'>C 2min</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Denature 98</span><span
 +
style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-mso-fareast-theme-font:
 +
minor-fareast;mso-hansi-theme-font:minor-latin'>&#730;</span><span
 +
lang=EN-US style='font-size:11.0pt'>C 10sec</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Annealing 51</span><span
 +
style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-mso-fareast-theme-font:
 +
minor-fareast;mso-hansi-theme-font:minor-latin'>&#730;</span><span
 +
lang=EN-US style='font-size:11.0pt'>C 30sec</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Extension 68</span><span
 +
style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-mso-fareast-theme-font:
 +
minor-fareast;mso-hansi-theme-font:minor-latin'>&#730;</span><span
 +
lang=EN-US style='font-size:11.0pt'>C 1min30sec</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>(30 cycles)</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 style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 12<sup>th</sup></span><span
 +
lang=EN-US></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Electrophoresed
 +
PCR products in 1% <span class=SpellE>agarose</span> gel.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We detected the
 +
correct size of DNA band.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 13<sup>th</sup></span><sup><span
 +
lang=EN-US></span></sup></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Made competent
 +
cells</span><span lang=EN-US>.</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 style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 17<sup>th</sup></span><sup><span
 +
lang=EN-US></span></sup></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified the PCR
 +
product.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><sup><span
 +
lang=EN-US>&nbsp;</span></sup></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 20<sup>th</sup></span><span
 +
lang=EN-US></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Electrophoresed <span
 +
class=GramE><i style='mso-bidi-font-style:normal'>ATF1</i>(</span>prepared on
 +
6/17) in 1% <span class=SpellE>agarose</span> gel.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Many DNA fragments
 +
were detected (failed PCR).</span></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 21<sup>st</sup></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>PCR was carried
 +
out.</span></p>
 +
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
 +
mso-yfti-tbllook:1184;mso-padding-alt:0mm 5.4pt 0mm 5.4pt'>
 +
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  mso-border-alt:solid windowtext .5pt;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=51 valign=top style='width:51.4pt;border:solid windowtext 1.0pt;
 +
  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
 +
  solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>50&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:1'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>dNTP</span></span><span
 +
  lang=EN-US style='font-size:11.0pt'></span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:2'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:3'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><i style='mso-bidi-font-style:normal'><span lang=EN-US
 +
  style='font-size:11.0pt'>ATF1</span></i><span lang=EN-US style='font-size:
 +
  11.0pt'>(PCR product)</span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:4'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>30cycle
 +
(Annealing:52</span><span style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:
 +
"";mso-fareast-theme-font:minor-fareast;
 +
mso-hansi-theme-font:minor-latin'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C)</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>PCR products were
 +
electrophoresed in 1% <span class=SpellE>agarose</span> gel.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Many DNA fragments
 +
were detected (failed PCR).</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>PCR was carried
 +
out.</span></p>
 +
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
 +
mso-yfti-tbllook:1184;mso-padding-alt:0mm 5.4pt 0mm 5.4pt'>
 +
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  mso-border-alt:solid windowtext .5pt;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=51 valign=top style='width:51.4pt;border:solid windowtext 1.0pt;
 +
  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
 +
  solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>50&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:1'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>dNTP</span></span><span
 +
  lang=EN-US style='font-size:11.0pt'></span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:2'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:3'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><i style='mso-bidi-font-style:normal'><span lang=EN-US
 +
  style='font-size:11.0pt'>ATF1</span></i><span lang=EN-US style='font-size:
 +
  11.0pt'>(PCR product)</span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:4'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>30cycle
 +
(Annealing:53</span><span style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:
 +
"";mso-fareast-theme-font:minor-fareast;
 +
mso-hansi-theme-font:minor-latin'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C)</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 24<sup>th</sup></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>PCR products were
 +
electrophoresed in 1% <span class=SpellE>agarose</span> gel.</span></p>
 +
 +
<p class=MsoNormal><span class=GramE><span lang=EN-US style='font-size:11.0pt'>Isolated
 +
and purified <i style='mso-bidi-font-style:normal'>ATF1</i> from 1% <span
 +
class=SpellE>agarose</span> gel.</span></span><span lang=EN-US
 +
style='font-size:11.0pt'></span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR
 +
to amplify the <i style='mso-bidi-font-style:normal'>ATF1</i> gene.</span></p>
 +
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
 +
mso-yfti-tbllook:1184;mso-padding-alt:0mm 5.4pt 0mm 5.4pt'>
 +
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  mso-border-alt:solid windowtext .5pt;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=51 valign=top style='width:51.4pt;border:solid windowtext 1.0pt;
 +
  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
 +
  solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>50&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:1'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>dNTP</span></span><span
 +
  lang=EN-US style='font-size:11.0pt'></span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:2'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:3'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><i style='mso-bidi-font-style:normal'><span lang=EN-US
 +
  style='font-size:11.0pt'>ATF1</span></i><span lang=EN-US style='font-size:
 +
  11.0pt'>(PCR product)</span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:4'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>30cycle
 +
(Annealing:52</span><span style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:
 +
"";mso-fareast-theme-font:minor-fareast;
 +
mso-hansi-theme-font:minor-latin'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>C)</span><span style='font-size:11.0pt;font-family:
 +
"";mso-ascii-theme-font:minor-latin;
 +
mso-fareast-mso-fareast-theme-font:minor-fareast;
 +
mso-hansi-theme-font:minor-latin'>)</span><span
 +
lang=EN-US style='font-size:11.0pt'></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 style='mso-bidi-font-weight:normal'><sup><span
 +
lang=EN-US>&nbsp;</span></sup></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><sup><span
 +
lang=EN-US>&nbsp;</span></sup></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><sup><span
 +
lang=EN-US>&nbsp;</span></sup></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><sup><span
 +
lang=EN-US>&nbsp;</span></sup></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 27<sup>th</sup></span><span
 +
lang=EN-US></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>PCR products were
 +
electrophoresed in 1% <span class=SpellE>agarose</span> gel.</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Four DNA fragments
 +
were detected</span></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>&nbsp;</span></b></p>
 +
 +
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-size:11.0pt'>June 28<sup>th</sup></span></b></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR
 +
to amplify the ATF1 gene.</span></p>
 +
 +
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
 +
style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
 +
mso-yfti-tbllook:1184;mso-padding-alt:0mm 5.4pt 0mm 5.4pt'>
 +
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  mso-border-alt:solid windowtext .5pt;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=51 valign=top style='width:51.4pt;border:solid windowtext 1.0pt;
 +
  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
 +
  solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>50&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:1'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span class=SpellE><span lang=EN-US style='font-size:11.0pt'>dNTP</span></span><span
 +
  lang=EN-US style='font-size:11.0pt'></span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:2'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:3'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><i style='mso-bidi-font-style:normal'><span lang=EN-US
 +
  style='font-size:11.0pt'>ATF1</span></i><span lang=EN-US style='font-size:
 +
  11.0pt'>(PCR product)</span></p>
 +
  </td>
 +
  <td width=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:4'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
<tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes'>
 +
  <td width=120 valign=top style='width:120.45pt;border:solid windowtext 1.0pt;
 +
  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
 +
  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=51 valign=top style='width:51.4pt;border-top:none;border-left:none;
 +
  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 +
  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 +
  mso-border-alt:solid windowtext .5pt;padding:0mm 5.4pt 0mm 5.4pt'>
 +
  <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26.5&micro;L</span></p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>30cycle
 +
(Annealing:53</span><span style='font-size:11.0pt;
 +
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:
 +
"";mso-fareast-theme-font:minor-fareast;
 +
mso-hansi-theme-font:minor-latin'>&#730;</span><span lang=EN-US
 +
style='font-size:11.0pt'>Cand54</span><span style='font-size:11.0pt;font-family:
 +
"";mso-ascii-theme-font:minor-latin;
 +
mso-fareast-mso-fareast-theme-font:minor-fareast;
 +
mso-hansi-theme-font:minor-latin'>&#730;</span><span
 +
lang=EN-US style='font-size:11.0pt'>C)</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>&nbsp;</span></p>
</div>
</div>

Latest revision as of 08:54, 27 September 2013




ATF1

June 4th

Minipreped ATF1(prepared on 5/31).

 

 

June 6th

Performed PCR to amplify the ATF1 gene.

Buffer

50µL

dNTP

20µL

Primer Mix

1µL

Genome DNA

0.5µL

KOD-FX

2µL

H2O

26.5µL

Forward: TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC

Reverse: ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC

35cycle

 

 

June 7th

Electrophoresed PCR products in 1% agarose gel. No DNA was detected.

 

 

June 10th

Performed PCR to amplify the ATF1 gene.

Buffer

50µL

dNTP

20µL

Primer Mix

1µL

Genome DNA

0.5µL

KOD-FX

2µL

H2O

26.5µL

Predenature 94˚C 2min

Denature 98˚C 10sec

Annealing 51˚C 30sec

Extension 68˚C 1min30sec

(30 cycles)

 

 

 

 

June 11th

Electrophoresed PCR products in 1% agarose gel.

We detected the correct size of DNA band.

We performed PCR to amplify the ATF1 gene.

Buffer

50µL

dNTP

20µL

Primer Mix

1µL

ATF1(PCR product)

0.5µL

KOD-FX

2µL

H2O

26.5µL

Predenature 94˚C 2min

Denature 98˚C 10sec

Annealing 51˚C 30sec

Extension 68˚C 1min30sec

(30 cycles)

 

 

June 12th

Electrophoresed PCR products in 1% agarose gel.

We detected the correct size of DNA band.

 

 

June 13th

Made competent cells.

 

 

June 17th

Purified the PCR product.

 

 

June 20th

Electrophoresed ATF1(prepared on 6/17) in 1% agarose gel.

Many DNA fragments were detected (failed PCR).

 

 

 

 

 

June 21st

PCR was carried out.

Buffer

50µL

dNTP

20µL

Primer Mix

1µL

ATF1(PCR product)

0.5µL

KOD-FX

2µL

H2O

26.5µL

30cycle (Annealing:52˚C)

PCR products were electrophoresed in 1% agarose gel.

Many DNA fragments were detected (failed PCR).

PCR was carried out.

Buffer

50µL

dNTP

20µL

Primer Mix

1µL

ATF1(PCR product)

0.5µL

KOD-FX

2µL

H2O

26.5µL

30cycle (Annealing:53˚C)

 

 

June 24th

PCR products were electrophoresed in 1% agarose gel.

Isolated and purified ATF1 from 1% agarose gel.

We performed PCR to amplify the ATF1 gene.

Buffer

50µL

dNTP

20µL

Primer Mix

1µL

ATF1(PCR product)

0.5µL

KOD-FX

2µL

H2O

26.5µL

30cycle (Annealing:52˚C)

 

 

 

 

 

 

June 27th

PCR products were electrophoresed in 1% agarose gel.

Four DNA fragments were detected

 

 

June 28th

We performed PCR to amplify the ATF1 gene.

Buffer

50µL

dNTP

20µL

Primer Mix

1µL

ATF1(PCR product)

0.5µL

KOD-FX

2µL

H2O

26.5µL

30cycle (Annealing:53˚Cand54˚C)