832 pMiniT-VVTopo

6/23/2017

colony PCR  screening

6/24/17

Miniprep, digestion with EcoRV

1, 3: PCR using NEB primers

2, 4, PCR using pMiniT-seq-F and pMiniT-seq-R

7/5/2017

Fix clone using new primers

remove extra ATG PCR from 832 using these two primers 831-F ATCCATATGCGTGCCCTGTTCTATAAAG

831-R CTGTCTAGAGTCGACGGTACCAAG (CTTGGTACCGTCGACTCTAGACAG)

clone back into pMiniT 1.2

miniprep digestion, EcoRV 

Miniprep digestion with Nde1 + Xho1