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Team:KIT-Kyoto/Notebook/ATF1/july
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| - | < | + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July 1<sup>st</sup></span></b></p> |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR | |
| - | + | at 53</span><span style='font-size:11.0pt;font-family:"ＭＳ 明朝","serif"'>˚</span><span | |
| - | + | lang=EN-US style='font-size:11.0pt'>C and 54</span><span style='font-size:11.0pt; | |
| - | + | font-family:"ＭＳ 明朝","serif"'>˚</span><span lang=EN-US style='font-size:11.0pt'>C. | |
| - | + | (annealing temp.)</span></p> | |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied DNA | |
| - | + | sample to the agarose gel electrophoresis.</span></p> | |
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| - | applied DNA sample to the | + | |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>-53</span><span | |
| - | + | style='font-size:11.0pt;font-family:"ＭＳ 明朝","serif"'>˚</span><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>C ver.-</span></p> | |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We detected several | |
| - | + | bands.</span></p> | |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>-54</span><span | |
| - | + | style='font-size:11.0pt;font-family:"ＭＳ 明朝","serif"'>˚</span><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>C ver.-</span></p> | |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>The bands seems | |
| - | + | wrong size.</span></p> | |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1. We applied DNA | |
| - | + | sample to the blue gel electrophoresis and purified DNA.</span></p> | |
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| - | applied DNA sample to the blue gel electrophoresis.</ | + | |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> [Consequence]</span></p> | |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>The band appeared | |
| - | + | 1600bp.</span></p> | |
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| - | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2. We performed | |
| - | + | PCR at 51</span><span style='font-size:11.0pt;font-family:"ＭＳ 明朝","serif"'>˚</span><span | |
| - | + | lang=EN-US style='font-size:11.0pt'>C and 52</span><span style='font-size:11.0pt; | |
| - | + | font-family:"ＭＳ 明朝","serif"'>˚</span><span lang=EN-US style='font-size:11.0pt'>C. | |
| - | + | (annealing temp.)</span></p> | |
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| - | + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | |
| - | + | style='border-collapse:collapse;border:none'> | |
| - | + | <tr> | |
| - | + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | |
| - | + | padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>Buffer</span></p> | |
| - | + | </td> | |
| - | + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | |
| - | + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>50µL</span></p> | |
| - | + | </td> | |
| - | + | </tr> | |
| - | + | <tr> | |
| - | + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | |
| - | + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>dNTP</span></p> | |
| - | + | </td> | |
| - | + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | |
| - | + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | |
| - | + | padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>20µL</span></p> | |
| - | + | </td> | |
| - | + | </tr> | |
| - | + | <tr> | |
| - | + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | |
| - | + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>primer mix</span></p> | |
| - | + | </td> | |
| - | + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | |
| - | + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | |
| - | + | padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>1µL</span></p> | |
| - | + | </td> | |
| - | + | </tr> | |
| - | + | <tr> | |
| - | + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | |
| - | + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | |
| - | + | lang=EN-US style='font-size:11.0pt'>PCR product</span></p> | |
| - | + | </td> | |
| - | + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | |
| - | + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | |
| - | + | padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>4µL</span></p> | |
| - | + | </td> | |
| - | < | + | </tr> |
| - | + | <tr> | |
| - | < | + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; |
| - | + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | </ | + | style='font-size:11.0pt'>KOD-FX</span></p> |
| - | < | + | </td> |
| - | < | + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; |
| - | + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | |
| - | + | padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | < | + | style='font-size:11.0pt'>2µL</span></p> |
| - | + | </td> | |
| - | + | </tr> | |
| - | < | + | <tr> |
| - | + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | |
| - | + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | |
| - | + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | |
| - | < | + | style='font-size:11.0pt'>H<sub>2</sub>O</span></p> |
| - | + | </td> | |
| - | + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | |
| - | + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | |
| - | + | padding:0mm 5.4pt 0mm 5.4pt'> | |
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>23µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| - | + | <p class=MsoNormal><span lang=EN-US> </span></p> | |
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| - | + | <p class=MsoNormal><span lang=EN-US> </span></p> | |
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| - | + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July 2<sup>nd</sup></span></b></p> | |
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| - | + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | |
| - | + | style='font-size:11.0pt'>We applied DNA sample to the agarose gel | |
| - | + | electrophoresis.</span></p> | |
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| - | applied DNA sample to the | + | |
| - | + | ||
| - | + | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>[Consequence]</span></p> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>The band appeared at the correct size.</span></p> | ||
| + | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July 4<sup>th</sup></span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We purified DNA | ||
| + | sample (PCR product on July 1<sup>st</sup>.)</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested <i>ATF1</i> | ||
| + | with <i>Hin</i>dIII.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied DNA | ||
| + | sample to the agarose gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>[Consequence]</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>The band appeared | ||
| + | at the correct size.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR | ||
| + | for <i>ATF1</i>.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>50µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>dNTP</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>20µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>primer mix</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>1µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>PCR | ||
| + | product</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>4µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>KOD-FX</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>2µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=109 valign=top style='width:82.05pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US>23µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July5<sup>th</sup></span></b><b><span | ||
| + | style='font-size:14.0pt;font-family:"ＭＳ 明朝","serif"'>・</span></b><b><span | ||
| + | lang=EN-US style='font-size:14.0pt'>8<sup>th</sup></span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We purified <i>ATF1</i>.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested <i>ATF1 | ||
| + | </i>and pET-15b with <i>Xho</i>I.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We purified this <i>ATF1</i> and pET-15b.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July 9<sup>th</sup></span></b></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We digested <i>ATF1 </i>and pET-15b with <i>Bpu</i>1102I.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>ATF1</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>25µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>3µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>2µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>30µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>25µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>3µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>2µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>30µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We incubated at 37</span><span style='font-size:11.0pt; | ||
| + | font-family:"ＭＳ 明朝","serif"'>˚</span><span lang=EN-US style='font-size:11.0pt'>C(30min)</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We added 1uL BAP to vector.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We incubated at 37</span><span style='font-size:11.0pt; | ||
| + | font-family:"ＭＳ 明朝","serif"'>˚</span><span lang=EN-US style='font-size:11.0pt'>C(30min)</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We applied DNA sample to the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>[Consequence]</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>The band did not appear.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We performed PCR DNA sample (PCR product on July 1<sup>st</sup>.)</span></p> | ||
| + | |||
| + | <p class=MsoListParagraph style='margin-left:21.0pt'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoListParagraph style='margin-left:21.0pt'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <span lang=EN-US style='font-size:11.0pt;font-family:"Century","serif"'><br | ||
| + | clear=all style='page-break-before:always'> | ||
| + | </span> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July 11th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We purified <i>ATF1</i></span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We digested <i>ATF1 </i>with <i>Xho</i>I.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>ATF1</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>25</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Xho</span></i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>2</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Buffer(10xH)</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>3</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>30</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>88</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Xho</span></i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>2</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Buffer(10xH)</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>10</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>100</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We applied DNA sample to the agarose gel | ||
| + | electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We purified DNA sample.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We digested <i>ATF1 </i>and Vector<i> </i>with <i>Bpu</i>1102I.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>ATF1</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>25µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>3µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>2µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>30µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoListParagraph align=left style='margin-left:21.0pt;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>34µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>2µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>4µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph align=left style='margin-left:0mm;text-align:left'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>40µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We applied DNA sample to the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>[result]</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Failed</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July 12<sup>th</sup></span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR | ||
| + | at 51</span><span style='font-size:11.0pt;font-family:"ＭＳ 明朝","serif"'>˚</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>C and 48</span><span style='font-size:11.0pt; | ||
| + | font-family:"ＭＳ 明朝","serif"'>˚</span><span lang=EN-US style='font-size:11.0pt'>C</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>50</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>dNTP</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>20</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Primer mix</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>genome</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>0.5</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>KOD-FX</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>26.5</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied DNA | ||
| + | sample to the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>[result] Failed</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR | ||
| + | at 51</span><span style='font-size:11.0pt;font-family:"ＭＳ 明朝","serif"'>˚</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>C</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>50</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>dNTP</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>20</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Primer mix</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>ATF1plasmid</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>0.5</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>KOD-FX</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>26.5</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July 17<sup>th</sup></span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied DNA | ||
| + | sample to the agarose gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>50</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>dNTP</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>20</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Primer mix</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Genome DNA</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>0.5</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>KOD-FX</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>26.5</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We extracted | ||
| + | sample DNA for gel</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied DNA | ||
| + | sample to the agarose gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>50</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>dNTP</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>20</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Primer mix</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Genome DNA</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>0.5</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>KOD-FX</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>4</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=116 valign=top style='width:86.7pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>25</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July 18<sup>th</sup></span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We extracted | ||
| + | sample DNA for gel</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>50</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>dNTP</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>20</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Primer mix</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>ATF1</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>KOD-FX</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=100 valign=top style='width:75.25pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=67 valign=top style='width:50.0pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoListParagraph style='margin-left:0mm'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>25</span><span lang=EN-US style='font-size:11.0pt'>µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied DNA | ||
| + | sample to the agarose gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR | ||
| + | to use KOD-FXNeo and KOD-FX</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied DNA | ||
| + | sample to the agarose gel electrophoresis.</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 align=left style='text-align:left'><span lang=EN-US> </span></p> | ||
| + | |||
| + | <span lang=EN-US style='font-size:10.5pt;font-family:"Century","serif"'><br | ||
| + | clear=all style='page-break-before:always'> | ||
| + | </span> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>July 19<sup>th</sup></span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We purified PCR | ||
| + | products.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested<i> </i>sample | ||
| + | DNA<i> </i>with<i> Hind</i>III.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested<i> </i>pet-15b | ||
| + | and <i>ATF1</i> with <i>Xho</i>I.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>25µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer(10xH)</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>30µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>34µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer(10xH)</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>4µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=118 valign=top style='width:88.65pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>40µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We purified them.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We digested<i> </i>pET-15b | ||
| + | and <i>ATF1</i> with <i>Bpu</i>1102I.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>25µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>30µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='margin-left:42.0pt;border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>25µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=93 valign=top style='width:69.8pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>total</span></p> | ||
| + | </td> | ||
| + | <td width=55 valign=top style='width:41.2pt;border-top:none;border-left:none; | ||
| + | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>30µL</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We applied DNA | ||
| + | sample to the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
</div> | </div> | ||
| + | |||
</div> | </div> | ||
| - | |||
Latest revision as of 09:16, 27 September 2013
ATF1
July 1st
We performed PCR at 53˚C and 54˚C. (annealing temp.)
We applied DNA sample to the agarose gel electrophoresis.
-53˚C ver.-
We detected several bands.
-54˚C ver.-
The bands seems wrong size.
1. We applied DNA sample to the blue gel electrophoresis and purified DNA.
[Consequence]
The band appeared 1600bp.
2. We performed PCR at 51˚C and 52˚C. (annealing temp.)
|
Buffer |
50µL |
|
dNTP |
20µL |
|
primer mix |
1µL |
|
PCR product |
4µL |
|
KOD-FX |
2µL |
|
H2O |
23µL |
July 2nd
We applied DNA sample to the agarose gel electrophoresis.
[Consequence]
The band appeared at the correct size.
July 4th
We purified DNA sample (PCR product on July 1st.)
We digested ATF1 with HindIII.
We applied DNA sample to the agarose gel electrophoresis.
[Consequence]
The band appeared at the correct size.
We performed PCR for ATF1.
|
Buffer |
50µL |
|
dNTP |
20µL |
|
primer mix |
1µL |
|
PCR product |
4µL |
|
KOD-FX |
2µL |
|
H2O |
23µL |
July5th・8th
We purified ATF1.
We digested ATF1 and pET-15b with XhoI.
We purified this ATF1 and pET-15b.
July 9th
We digested ATF1 and pET-15b with Bpu1102I.
|
ATF1 |
25µL |
|
Buffer |
3µL |
|
Bpu1102I |
2µL |
|
total |
30µL |
|
pET-15b |
25µL |
|
Buffer |
3µL |
|
Bpu1102I |
2µL |
|
total |
30µL |
We incubated at 37˚C(30min)
We added 1uL BAP to vector.
We incubated at 37˚C(30min)
We applied DNA sample to the blue gel electrophoresis.
[Consequence]
The band did not appear.
We performed PCR DNA sample (PCR product on July 1st.)
July 11th
We purified ATF1
We digested ATF1 with XhoI.
|
ATF1 |
25µL |
|
XhoI |
2µL |
|
Buffer(10xH) |
3µL |
|
Total |
30µL |
|
pET-15b |
88µL |
|
XhoI |
2µL |
|
Buffer(10xH) |
10µL |
|
total |
100µL |
We applied DNA sample to the agarose gel electrophoresis.
We purified DNA sample.
We digested ATF1 and Vector with Bpu1102I.
|
ATF1 |
25µL |
|
Buffer |
3µL |
|
Bpu1102I |
2µL |
|
total |
30µL |
|
pET-15b |
34µL |
|
Buffer |
2µL |
|
Bpu1102I |
4µL |
|
total |
40µL |
We applied DNA sample to the blue gel electrophoresis.
[result]
Failed
July 12th
We performed PCR at 51˚C and 48˚C
|
Buffer |
50µL |
|
dNTP |
20µL |
|
Primer mix |
1µL |
|
genome |
0.5µL |
|
KOD-FX |
2µL |
|
H2O |
26.5µL |
We applied DNA sample to the blue gel electrophoresis.
[result] Failed
We performed PCR at 51˚C
|
Buffer |
50µL |
|
dNTP |
20µL |
|
Primer mix |
1µL |
|
ATF1plasmid |
0.5µL |
|
KOD-FX |
2µL |
|
H2O |
26.5µL |
July 17th
We applied DNA sample to the agarose gel electrophoresis.
We performed PCR
|
Buffer |
50µL |
|
dNTP |
20µL |
|
Primer mix |
1µL |
|
Genome DNA |
0.5µL |
|
KOD-FX |
2µL |
|
H2O |
26.5µL |
We extracted sample DNA for gel
We applied DNA sample to the agarose gel electrophoresis.
We performed PCR.
|
Buffer |
50µL |
|
dNTP |
20µL |
|
Primer mix |
1µL |
|
Genome DNA |
0.5µL |
|
KOD-FX |
4µL |
|
H2O |
25µL |
July 18th
We extracted sample DNA for gel
We performed PCR.
|
Buffer |
50µL |
|
dNTP |
20µL |
|
Primer mix |
1µL |
|
ATF1 |
2µL |
|
KOD-FX |
2µL |
|
H2O |
25µL |
We applied DNA sample to the agarose gel electrophoresis.
We performed PCR to use KOD-FXNeo and KOD-FX
We applied DNA sample to the agarose gel electrophoresis.
July 19th
We purified PCR products.
We digested sample DNA with HindIII.
We digested pet-15b and ATF1 with XhoI.
|
ATF1 |
25µL |
|
XhoI |
2µL |
|
Buffer(10xH) |
3µL |
|
total |
30µL |
|
pET-15b |
34µL |
|
XhoI |
2µL |
|
Buffer(10xH) |
4µL |
|
total |
40µL |
We purified them.
We digested pET-15b and ATF1 with Bpu1102I.
|
pET-15b |
25µL |
|
Buffer |
3µL |
|
Bpu1102I |
2µL |
|
total |
30µL |
|
ATF1 |
25µL |
|
Buffer |
3µL |
|
Bpu1102I |
2µL |
|
total |
30µL |
We applied DNA sample to the blue gel electrophoresis.
