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<p>X35</p>
<p> </p>
<p> [[File:14.png|400px]]</p>
<p>-> Can see the insert(300bp) in both 54,56,58 temperature gradient!</p>
<p> </p>
<p> (2)</p>
<p><T vector ligation></p>
<p>Insert DNA mass: 8.607ng(3:1)</p>
<p>2X Rapid ligation: 5ul</p>
<p>T vector: 0.5ul(25ng)</p>
<p>PCR product: 1ul(8.7ng)</p>
<p>D.W: 2.5ul</p>
<p>T4 DNA Ligase: 1ul</p>
<p>----------------------------</p>
<p>Total: 10ul</p>
<p>RT 1 hour incubation</p>
<p>Then, Transformation</p>
<p> </p>
<p> </p>
<p> <Colony PCR></p>
<p>-> Check whether insert base pairs is inserted in T vector well</p>
<p>T.D.W : 14.9</p>
<p>10X Buffer : 2</p>
<p>M13 primer Forward: 0.5</p>
<p>M13 primer Reverse: 0.5</p>
<p>2.5mM dNTP: 1.6</p>
<p>XL-Taq polymerase: 0.5</p>
<p> </p>
<p> ---------------------------------</p>
<p> </p>
<p> Can check on 3,5 well(T vector 200bp+ 346bp = 500~600bp)</p>
<p> </p>
