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Team:KIT-Kyoto/Notebook/ATF1/august
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| + | <h2>ATF1</h2> | ||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 6th</span></b></p> | ||
| - | < | + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>The <i>ATF1 </i>gene |
| - | + | was purified (prepared on 7/19)</span></p> | |
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| - | < | + | |
| - | was purified (prepared on 7/19)</ | + | |
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| - | + | <p class=MsoNormal align=left style='text-align:left;text-autospace:none'><span | |
| - | + | lang=EN-US style='font-size:11.0pt'>Digested pET-15b with <i>Xho</i>I at 37</span><span | |
| - | + | lang=EN-US style='font-size:11.0pt;font-family:メイリオ'> </span><span | |
| - | + | style='font-size:11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span | |
| - | + | lang=EN-US style='font-size:11.0pt'> for 90 minutes.</span></p> | |
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| - | + | ||
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| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>88</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>100</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified pET-15b DNA | ||
| + | was dissolved in 34</span><span lang=EN-US style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L of H<sub>2</sub>O. </span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left;text-autospace:none'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Digested pET-15b with <i>Bpu</i>1102I at 37</span><span | ||
| + | lang=EN-US style='font-size:11.0pt;font-family:メイリオ'> </span><span | ||
| + | style='font-size:11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span | ||
| + | lang=EN-US style='font-size:11.0pt'> for 90 minutes.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>34</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>4</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>40</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left;text-autospace:none'><span | ||
| + | lang=EN-US style='font-size:11.0pt'>Added 1uL of BAP to the solution and | ||
| + | incubated it at 37</span><span lang=EN-US style='font-size:11.0pt;font-family: | ||
| + | メイリオ'> </span><span style='font-size:11.0pt;font-family:メイリオ'>˚<span | ||
| + | lang=EN-US>C </span></span><span lang=EN-US style='font-size:11.0pt'>for 30 | ||
| + | minutes.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied <i>ATF1</i> | ||
| + | and pET-15b to the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Results: No band | ||
| + | was detected.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Cultivated pET-15b | ||
| + | in 3mL LB medium with ampicillin overnight.</span></p> | ||
| + | |||
| + | <span lang=EN-US style='font-size:11.0pt;font-family:"Century","serif"'><br | ||
| + | clear=all style='page-break-before:always'> | ||
| + | </span> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 7th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Minipreped pET-15b | ||
| + | DNA.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b | ||
| + | with <i>Xho</i>I at 37</span><span style='font-size:11.0pt;font-family:メイリオ'>˚<span | ||
| + | lang=EN-US>C</span></span><span lang=EN-US style='font-size:11.0pt'> for 3 | ||
| + | hours.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>88</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>100</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified it and dissolved | ||
| + | in 26</span><span lang=EN-US style='font-size:11.0pt'>µ</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>L of H<sub>2</sub>O.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b | ||
| + | with <i>Bpu</i>1102I at 37</span><span style='font-size:11.0pt;font-family: | ||
| + | メイリオ'>˚<span lang=EN-US>C</span></span><span lang=EN-US style='font-size:11.0pt'> | ||
| + | for 3 hours.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>30</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Added 1</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L of BAP to this solution and incubated it at 37</span><span | ||
| + | style='font-size:11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span | ||
| + | lang=EN-US style='font-size:11.0pt'> for 30 minutes.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied <i>ATF1</i> | ||
| + | and pET-15b to the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <span lang=EN-US style='font-size:11.0pt;font-family:"Century","serif"'><br | ||
| + | clear=all style='page-break-before:always'> | ||
| + | </span> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 8th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b | ||
| + | with <i>Xho</i>I at 37</span><span style='font-size:11.0pt;font-family:メイリオ'>˚<span | ||
| + | lang=EN-US>C</span></span><span lang=EN-US style='font-size:11.0pt'> overnight.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>89</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>100</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </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'>August 9th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified pET-15b (prepared | ||
| + | on 8/8) and added 26ul of H<sub>2</sub>O.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b | ||
| + | with <i>Bpu</i>1102I at 37</span><span style='font-size:11.0pt;font-family: | ||
| + | メイリオ'>˚<span lang=EN-US>C</span></span><span lang=EN-US style='font-size:11.0pt'> | ||
| + | for 3 hours.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>30</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Added 1</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L of BAP to this solution and incubate it at 37</span><span | ||
| + | style='font-size:11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span | ||
| + | lang=EN-US style='font-size:11.0pt'> for 30 minutes.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied pET-15b to | ||
| + | the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Isolated and | ||
| + | Purified it.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligation pET-15b | ||
| + | and <i>ATF1 </i>(prepared 7/19,8/6) at room temperature for 15 minutes.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>5</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>5</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligation MIX</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>The <i>ATF1 </i>into | ||
| + | pET-15b was transformed into <i>E.coli </i>cells.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Cultivated | ||
| + | transformants on LB plate with ampicillin at37</span><span style='font-size: | ||
| + | 11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span lang=EN-US | ||
| + | style='font-size:11.0pt'> overnight.</span></p> | ||
| + | |||
| + | <span lang=EN-US style='font-size:11.0pt;font-family:"Century","serif"'><br | ||
| + | clear=all style='page-break-before:always'> | ||
| + | </span> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 10th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked 48 colonies | ||
| + | of <i>ATF1 </i>transformants.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 12th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Checked the | ||
| + | colonies by colony cracking.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Picked up the | ||
| + | appropriate colonies and cultured in the LB in 3mL ampicillin(+) medium at 37</span><span | ||
| + | style='font-size:11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span | ||
| + | lang=EN-US style='font-size:11.0pt'> for 4 hours.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Miniprepped plasmid | ||
| + | DNA.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 13th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested plasmid | ||
| + | DNA (prepared on 8/12) with<i> Hin</i>dIII at 37</span><span style='font-size: | ||
| + | 11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span lang=EN-US | ||
| + | style='font-size:11.0pt'> for 90 minutes.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Plasmid DNA</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>5</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>12</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Hin</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>dIII</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied pET-15b to | ||
| + | the agarose gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested plasmid | ||
| + | DNA (prepared on 8/12) with <i>Hin</i>dIII at 37</span><span style='font-size: | ||
| + | 11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span lang=EN-US | ||
| + | style='font-size:11.0pt'> for 90 minutes.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Plasmid DNA</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>7.25</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Hin</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>dIII</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.75</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Total</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied pET-15b to | ||
| + | the agarose gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Again, picked up the | ||
| + | appropriate colonies and cultured in the 3mL LB medium with ampicillin at 37</span><span | ||
| + | style='font-size:11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <b><span lang=EN-US style='font-size:14.0pt;font-family:"Century","serif"'><br | ||
| + | clear=all style='page-break-before:always'> | ||
| + | </span></b> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US | ||
| + | style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 26th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>We performed PCR | ||
| + | to amplify the <i>ATF1 </i>gene.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>50</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>dNTP</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>20</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Primer mix</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Genome DNA</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>0.5</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>KOD-FX</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>26.5</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:14.0pt'>August 27th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified PCR | ||
| + | products (prepared on 8/26) and added 30</span><span lang=EN-US | ||
| + | style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size:11.0pt'>L of | ||
| + | H<sub>2</sub>O.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested <i>ATF1</i> | ||
| + | with <i>Hin</i>dIII.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>5</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Hin</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>dIII</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>11</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b | ||
| + | and <i>ATF1</i> with <i>Xho</i>I.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>25</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>100</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>11.3</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified it and | ||
| + | added 25</span><span lang=EN-US style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L of H<sub>2</sub>O.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested pET-15b | ||
| + | and <i>ATF1 </i>with <i>Bpu</i>1102I.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>25</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>25</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>2</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied pET-15b | ||
| + | and <i>ATF1</i> to the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified PCR | ||
| + | products.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><b><span lang=EN-US style='font-size:11.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US | ||
| + | style='font-size:14.0pt'>August 28th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Digested <i>ATF1 </i>(prepared on 8/27) with <i>Eco</i>RI.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>5</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Eco</span></i><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>RI</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Applied it to the agarose gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Digested <i>ATF1 </i>(prepared on 8/27) and pET-15b | ||
| + | with <i>Xho</i>I.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>25</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>3</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Xho</span></i><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>100</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>11.3</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Xho</span></i><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US | ||
| + | style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US | ||
| + | style='font-size:14.0pt'> </span></b></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US | ||
| + | style='font-size:14.0pt'>August 29th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Applied <i>ATF1 </i>and pET-15b to the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Ligated it.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><i><span lang=EN-US | ||
| + | style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>10</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>10</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Ligation MIX</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>10</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Carried out transformation.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><b><span lang=EN-US | ||
| + | style='font-size:14.0pt'>August 30th</span></b></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Checked the colonies by colony cracking.</span></p> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>We performed PCR to amplify the <i>ATF1</i> gene.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Buffer</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>50</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>dNTP</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>20</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Primer mix</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>1</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>Genome DNA</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>0.5</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>KOD-FX</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>2</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>H<sub>2</sub>O</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'>26.5</span><span lang=ja style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal align=left style='text-align:left'><span lang=EN-US | ||
| + | style='font-size:11.0pt'> </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><span lang=EN-US style='font-size:14.0pt'>August 31<sup>st</sup></span></b></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified PCR | ||
| + | products and dissolved 30</span><span lang=EN-US style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L of H<sub>2</sub>O.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Purified pET-15b | ||
| + | and added 33</span><span lang=EN-US style='font-size:11.0pt'>µ</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>L of H<sub>2</sub>O.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Digested <i>ATF1</i> | ||
| + | and pET-15b with <i>Xho</i>I and <i>Bpu</i>1102I.</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NEB Buffer 2</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>3</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>24</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>NEB Buffer 2</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>4</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>33</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>BSA</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Xho</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>Bpu</span></i><span | ||
| + | lang=EN-US style='font-size:11.0pt'>1102I</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>1</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Added 1</span><span | ||
| + | lang=EN-US style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L of BAP to pET-15b and incubated them at 37</span><span | ||
| + | style='font-size:11.0pt;font-family:メイリオ'>˚<span lang=EN-US>C</span></span><span | ||
| + | lang=EN-US style='font-size:11.0pt'> for 30 minutes.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Applied pET-15b to | ||
| + | the blue gel electrophoresis.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Isolated and purified | ||
| + | it.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligation of | ||
| + | pET-15b with <i>ATF1</i> (prep).</span></p> | ||
| + | |||
| + | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;border:none'> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><i><span lang=EN-US style='font-size:11.0pt'>ATF1</span></i></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-left:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>pET-15b</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td width=162 valign=top style='width:121.45pt;border:solid windowtext 1.0pt; | ||
| + | border-top:none;padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Ligation | ||
| + | MIX</span></p> | ||
| + | </td> | ||
| + | <td width=162 valign=top style='width:121.45pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | ||
| + | padding:0mm 5.4pt 0mm 5.4pt'> | ||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>10</span><span | ||
| + | lang=ja style='font-size:11.0pt'>µ</span><span lang=EN-US style='font-size: | ||
| + | 11.0pt'>L</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'>Transformed <i>ATF1</i> | ||
| + | into pET-15b <i>E. coli</i> cells.</span></p> | ||
| + | |||
| + | <p class=MsoNormal><span lang=EN-US style='font-size:11.0pt'> </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> | ||
</div> | </div> | ||
Latest revision as of 09:18, 27 September 2013
ATF1
August 6th
The ATF1 gene was purified (prepared on 7/19)
Digested pET-15b with XhoI at 37 ˚C for 90 minutes.
|
pET-15b |
88µL |
|
XhoI Buffer |
10µL |
|
XhoI |
2µL |
|
Total |
100µL |
Purified pET-15b DNA was dissolved in 34µL of H2O.
Digested pET-15b with Bpu1102I at 37 ˚C for 90 minutes.
|
pET-15b |
34µL |
|
Bpu1102I Buffer |
4µL |
|
Bpu1102I |
2µL |
|
Total |
40µL |
Added 1uL of BAP to the solution and incubated it at 37 ˚C for 30 minutes.
Applied ATF1 and pET-15b to the blue gel electrophoresis.
Results: No band was detected.
Cultivated pET-15b in 3mL LB medium with ampicillin overnight.
August 7th
Minipreped pET-15b DNA.
Digested pET-15b with XhoI at 37˚C for 3 hours.
|
pET-15b |
88µL |
|
XhoI Buffer |
10µL |
|
XhoI |
2µL |
|
Total |
100µL |
Purified it and dissolved in 26µL of H2O.
Digested pET-15b with Bpu1102I at 37˚C for 3 hours.
|
pET-15b |
26µL |
|
Bpu1102I Buffer |
3µL |
|
Bpu1102I |
1µL |
|
Total |
30µL |
Added 1µL of BAP to this solution and incubated it at 37˚C for 30 minutes.
Applied ATF1 and pET-15b to the blue gel electrophoresis.
August 8th
Digested pET-15b with XhoI at 37˚C overnight.
|
pET-15b |
89µL |
|
XhoI Buffer |
10µL |
|
XhoI |
1µL |
|
Total |
100µL |
August 9th
Purified pET-15b (prepared on 8/8) and added 26ul of H2O.
Digested pET-15b with Bpu1102I at 37˚C for 3 hours.
|
pET-15b |
26 |
|
Bpu1102I Buffer |
3µL |
|
Bpu1102I |
1µL |
|
Total |
30µL |
Added 1µL of BAP to this solution and incubate it at 37˚C for 30 minutes.
Applied pET-15b to the blue gel electrophoresis.
Isolated and Purified it.
Ligation pET-15b and ATF1 (prepared 7/19,8/6) at room temperature for 15 minutes.
|
ATF1 |
5µL |
|
pET-15b |
5µL |
|
Ligation MIX |
10µL |
|
Total |
20µL |
The ATF1 into pET-15b was transformed into E.coli cells.
Cultivated transformants on LB plate with ampicillin at37˚C overnight.
August 10th
Picked 48 colonies of ATF1 transformants.
August 12th
Checked the colonies by colony cracking.
Picked up the appropriate colonies and cultured in the LB in 3mL ampicillin(+) medium at 37˚C for 4 hours.
Miniprepped plasmid DNA.
August 13th
Digested plasmid DNA (prepared on 8/12) with HindIII at 37˚C for 90 minutes.
|
Plasmid DNA |
5µL |
|
Buffer |
2µL |
|
H2O |
12µL |
|
HindIII |
1µL |
|
Total |
20µL |
Applied pET-15b to the agarose gel electrophoresis.
Digested plasmid DNA (prepared on 8/12) with HindIII at 37˚C for 90 minutes.
|
Plasmid DNA |
10µL |
|
Buffer |
2µL |
|
H2O |
7.25µL |
|
HindIII |
0.75µL |
|
Total |
20µL |
Applied pET-15b to the agarose gel electrophoresis.
Again, picked up the appropriate colonies and cultured in the 3mL LB medium with ampicillin at 37˚C.
August 26th
We performed PCR to amplify the ATF1 gene.
|
Buffer |
50µL |
|
dNTP |
20µL |
|
Primer mix |
1µL |
|
Genome DNA |
0.5µL |
|
KOD-FX |
2µL |
|
H2O |
26.5µL |
August 27th
Purified PCR products (prepared on 8/26) and added 30µL of H2O.
Digested ATF1 with HindIII.
|
ATF1 |
5µL |
|
Buffer |
2µL |
|
HindIII |
2µL |
|
H2O |
11µL |
Digested pET-15b and ATF1 with XhoI.
|
ATF1 |
25µL |
|
Buffer |
3µL |
|
XhoI |
2µL |
|
pET-15b |
100µL |
|
Buffer |
11.3µL |
|
XhoI |
2µL |
Purified it and added 25µL of H2O.
Digested pET-15b and ATF1 with Bpu1102I.
|
ATF1 |
25µL |
|
Buffer |
3µL |
|
Bpu1102I |
2µL |
|
pET-15b |
25µL |
|
Buffer |
3µL |
|
Bpu1102I |
2µL |
Applied pET-15b and ATF1 to the blue gel electrophoresis.
Purified PCR products.
August 28th
Digested ATF1 (prepared on 8/27) with EcoRI.
|
ATF1 |
5µL |
|
Buffer |
1µL |
|
EcoRI |
2µL |
|
H2O |
2µL |
Applied it to the agarose gel electrophoresis.
Digested ATF1 (prepared on 8/27) and pET-15b with XhoI.
|
ATF1 |
25µL |
|
Buffer |
3µL |
|
XhoI |
2µL |
|
pET-15b |
100µL |
|
Buffer |
11.3µL |
|
XhoI |
2µL |
August 29th
Applied ATF1 and pET-15b to the blue gel electrophoresis.
Ligated it.
|
ATF1 |
10µL |
|
pET-15b |
10µL |
|
Ligation MIX |
10µL |
Carried out transformation.
August 30th
Checked the colonies by colony cracking.
We performed PCR to amplify the ATF1 gene.
|
Buffer |
50µL |
|
dNTP |
20µL |
|
Primer mix |
1µL |
|
Genome DNA |
0.5µL |
|
KOD-FX |
2µL |
|
H2O |
26.5µL |
August 31st
Purified PCR products and dissolved 30µL of H2O.
Purified pET-15b and added 33µL of H2O.
Digested ATF1 and pET-15b with XhoI and Bpu1102I.
|
NEB Buffer 2 |
3µL |
|
ATF1 |
24µL |
|
BSA |
1µL |
|
XhoI |
1µL |
|
Bpu1102I |
1µL |
|
NEB Buffer 2 |
4µL |
|
pET-15b |
33µL |
|
BSA |
1µL |
|
XhoI |
1µL |
|
Bpu1102I |
1µL |
Added 1µL of BAP to pET-15b and incubated them at 37˚C for 30 minutes.
Applied pET-15b to the blue gel electrophoresis.
Isolated and purified it.
Ligation of pET-15b with ATF1 (prep).
|
ATF1 |
10µL |
|
pET-15b |
10µL |
|
Ligation MIX |
10µL |
Transformed ATF1 into pET-15b E. coli cells.
