self-ligating vector - I need help/advice! (Mar/23/2005 )
Hi, I have been trying to make a few libraries using pSMART vector for a while now and I keep getting high amounts of vector background. My usual routine is to end repair and phosphorylate my fragments, size select on a gel, purify from the gel (TAE) and ligate to the vector provided by lucigen, but I am still getting an unbelievable amount od vector that appears to be self ligating or some kind of nucleotide contamination. Has anyone else used this kit? had this problem? any ideas? I am at the end of my rope.
-jodi-
you can see this topic
http://www.protocol-online.org/forums/inde...hl=dephosphoryl
-fred_33-