Editing 1297 pET-6H-VPMLK-TAT-NLS (section)

=== Cloning<br>  ===

Date:&nbsp;August 13, 2012\ 

Gene synthesis 

Purify PCR product, see lane 2, (113 bp, need to add 10% isopropanol becasue the fragment is very small). 

[[Image:1297 0813 Amp.JPG]] 

Digest PCR product with RE Nco1 + Nde1.<br>Digest vector 1128 pDB.His.PfuMBP-(tev) with Nco1 + Nde1.<br>Ligate together overnight and transform<br> 

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Date:&nbsp;August 14, 2012 

Insert disappeared after digestion, need to reamplify, see lane 1 

[[Image:0814 Amp.JPG]] 

Date:&nbsp;August 15, 2012<br> 

[[Image:1297 0815 Digestion.JPG]]<br> 

Digest PCR product with RE Nco1 + Nde1 (lane 1)<br>Digest vector pDB.His.MBP, V14 with Nco1 + Nde1 (lane 2)<br>Ligate together and transform<br>