Difference between revisions of "Team:Tianjin/Note/6803"

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<p><li><i>19</i> amplification at 65.0°C with 19.rev/fwd primes</li>
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<p>aaa
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      <li>PCR worked, positive control worked, no amplification of <i>19</i></li>
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<li><i>15</i> amplification at 65.0°C with 15.rev/fwd primes</li>
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      <li>PCR worked, positive control worked, no amplification of <i>15</i></li>
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<li>A product length of 900 bp was expected. The gel shows that most plasmids seem to be correct.</li>
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<li>This result was confirmed by sequencing.</li>
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<li>The fragments of <i>19</i> and <i>15</i> were purified with PCR Purification Kit.</li>
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<b>Ligation of <i>15</i> with <i>19</i><b><br/>
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<li>Fragment of <i>19</i> was phosphorylated and then purified with DNA Purification Kit.</li>
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<li><i>19</i> was ligated into T vectors via TA clone and transformed into E.coli via heat shock.</li>
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<li>A colony PCR was performed with five colonies</li>
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  <li>Gel electrophoresis showed that 5 colonies were positive for insertion of <i>19</i>.</li>
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<li>Two of these colonies containing <i>19</i> were used to inoculate overnight cultures.</li>
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  <li><i>15</i> gene fragment was phosphorylated.</li>
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<li>Plasmids containing T vector with <i>19</i>(pT-19)were isolated using a miniprep kit.</li>
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<li>Mono-restriction digest of pT-19 with stu I </li>
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<li>The enzyme-digested product was dephosphorylation.</li>
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<li>Dephosphorylated plasmid and phosphorylated gene <i>15</i> were connected.</li>
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  <li>Ligation product was transformed into E.coli via heat shock.</li>
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<li>A colony PCR was performed with twelve colonies.</li>
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      <li>Gel electrophoresis showed that 2 colonies were positive for connecting the plasmid and gene fragment.</li>
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      <li>These two colonies were used to inoculate overnight cultures.</li>
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<li>Plasmids with correct sequence of  <i>19-15</i>  were isolated using a miniprep kit.</li>
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<li>Mono-restriction digest of pT-19-15 with Nru I</li>
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<li>The enzyme-digested product was dephosphorylation.</li>
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Revision as of 14:08, 28 September 2016

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火影忍者疾风传

终于来到喀纳斯——中国最西北的角落，此次西北之行可到达的最远处。在这个纬度高达 48 度的地方，山坡上的植被更加茂密；太阳下落得很慢，阳光以一个非常低的角度横扫过来，跨过群山，透过树木，在地上都投出长长的影子，非常好看。

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火影忍者十分钟疾风传

告别禾木乡，继续向喀纳斯方向前行，途中经过冲乎尔乡。这里是一个天然牧场，坐落在小盆地，群山环抱，乡民主要以放牧为生。果然，不时就能看见牧羊人赶着几百头“阿勒泰大尾羊”在公路上浩浩荡荡地走着，一路扬起滚滚烟尘，甚是壮观。
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北疆树林

继续在铁热克提附近沿着蜿蜒的小道行摄，不知不觉间便绕到这座高山的另一侧。这里的杨树长得更高耸茂密，西斜的阳光恰到好处地将树梢照得金黄透亮，一道道树影投射在绒毛地毯般的甸上，构成近乎完美的光影，令人心旷神怡，不舍离。
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Notice: This page is currently under construction. Contents in this page are temporaory and will be modified several times before the final release.     — 2016 iGEM Team Tianjin