205 pET28-MBP-TurboTEV

Cloning of pET28-MBP-TurboTEV

By Paul

4/17/2012:

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

4/17/2012

digest vector 203 with Nde1+ EcoR1 o/n

digest PCR product with Nde1 + EcoR1 o/n

4/18/2012

1: vector 203, pET28-TurboTEV

2, 3: PCR product

gel purify, ligate and transform NEB DH5plah

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

Select 6 clone to do PCR screening

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

205 PCR Screening 4/20/2012

4/26/2012

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