2746

20200514 2746 pminiT2 NEBturbo miniprep EcoRI

2746 PCR

pET28-3XFlag-N miniprep digestion

PCR, 6/5/2020

ignore this 2696-pet28b vector. it failed

2746-pJet1.2 and subclone

20200616 2746 pJet NcoI-HF+XhoI in Cutsmart, 37oC, 1hour

20200617 2746-pJet2.1 digested with NcoI+XhoI. But two bands, one is between 1-2kb and the other one around 3kb, can be seen and cut for ligation with vector pET28b- seperately. Which one is the correct insert will be further determined.

20200617 2696/940/2696 all three vectors can provide pET28b-. Select 2696 for further subclone.

20200619 2746-pET28b clone PCR was performed. Primers used will produce 1385bp product. So the insert band between 1-2kb looks like the one we want.