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'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </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'>50µL</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'>20µL</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'>1µL</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'>0.5µL</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'>2µL</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'>26.5µL</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'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </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'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </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'>50µL</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'>20µL</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'>1µL</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'>0.5µL</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'>2µL</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'>26.5µL</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'>˚</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'>˚</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'>˚</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'>˚</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'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </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'>50µL</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'>20µL</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'>1µL</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'>0.5µL</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'>2µL</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'>26.5µL</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'>˚</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'>˚</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'>˚</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'>˚</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'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </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'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </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> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US> </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> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><sup><span | ||
+ | lang=EN-US> </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> </span></b></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US> </span></b></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US> </span></b></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US> </span></b></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US> </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'>50µL</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'>20µL</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'>1µL</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'>0.5µL</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'>2µL</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'>26.5µL</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'>˚</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'>50µL</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'>20µL</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'>1µL</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'>0.5µL</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'>2µL</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'>26.5µL</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'>˚</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'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US> </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'>50µL</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'>20µL</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'>1µL</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'>0.5µL</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'>2µL</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'>26.5µL</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'>˚</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'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><sup><span | ||
+ | lang=EN-US> </span></sup></b></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><sup><span | ||
+ | lang=EN-US> </span></sup></b></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><sup><span | ||
+ | lang=EN-US> </span></sup></b></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><sup><span | ||
+ | lang=EN-US> </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> </span></b></p> | ||
+ | |||
+ | <p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US> </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'>50µL</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'>20µL</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'>1µL</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'>0.5µL</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'>2µL</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'>26.5µL</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'>˚</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'>˚</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'> </span></p> | ||
+ | |||
+ | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </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)