Editing 205 pET28-MBP-TurboTEV

Cloning of pET28-MBP-TurboTEV<br> 

By Paul<br> 

4/17/2012: <br> 

PCR MBP from pDB.His.MBP<br>MBP-NdeI-F AGT CAC ATA TGA AAA CTG AAG AAG GTA AAC TG<br>MBP-EcoR1-R TCG ATG AAT TCA GTC TGC GCG TCT TTC AGG G<br> 

[[Image:1063 digestion 4 17 2012.jpg]]<br> 

4/17/2012<br> 

digest vector 203 with Nde1+ EcoR1 o/n<br> 

digest PCR product with Nde1 + EcoR1 o/n<br> 

4/18/2012<br> 

[[Image:205 digestion 4 18 2012.jpg]]<br> 

1: vector 203, pET28-TurboTEV<br> 

2, 3: PCR product 

gel purify, ligate and transform NEB DH5plah<br> 

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digest with Nde1+ EcoR1, clone into pET28-TurboTev 203<br> 

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Select 6 clone to do PCR screening<br> 

[[Image:1063 205 PCR Screening 4.19.2012.JPG]] 

Select 205-1,-2,-3 clones to mini-prep, digest 

[[Image:1063 205 digest 4.20.2012.JPG]] 

205 PCR Screening 4/20/2012 

[[Image:205 PCR Screening 4.20.2012.JPG]] 

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4/26/2012<br> 

[[Image:203i 1116v digest 4.26.2012.JPG]]<br> 

Select 4 clones to mini-prep<br>205 plasmids are digested by EcoR1 + Sal1<br> 

[[Image:205 digest 4.30.2012.JPG]]<br> 

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[[Category:Cloning]]