vector auto ligation problem - (Mar/23/2005 )
Hi guys, my subclone insert into pGEX4T2 vectors. I picked up some colonies and did mini pre of plasmids, the products gave me positive PCR test result, so I did not do double digestion. But expression result gave me only GST protein. Any advice? I wonder why the PCR test is not dependable.
-mhtprc-
how do you know that it is only the GST being expressed?
is your DNA fragment in frame with the GST? is there a premature stop codon?
I'd still the do the double digest on your minipreps just to see if it is consistent with your PCR result.
-george@CASE-