What are the diagnostic TRC pLKO.1 restriction digests?
BamHI digest gives 7032 band (linearizes the plasmid). BamHI/NdeI double digest gives 6238 and 794 bands. Note - BamHI is susceptible to Star Activity. If digests looks smeary, this could be the culprit. New Engand Biolabs website has some guidelines for Star Activity. http: //www. neb. com/nebecomm/default. asp