phosphatase question (library construction)

Alex Brands abbrands at
Mon Mar 10 13:39:41 EST 1997

I'm about to attempt construction of a cDNA library into a GAL4AD plasmid.
I have the mRNA, I have the kit, and I have cells that are 2X10^8/ug, so
I'm almost ready to go.  The problem is, I tried a ligation of the
prepared vector, and I got some background.  Not much, but some.  It was
prepared by cutting with EcoR1, CIP treatment, followed by gel
purification, organic extraction, precipitation.  

My question is this; If I use more CIP, I could reduce the background, but 
are there any possible detrimental effects of using too much CIP?
How about SIP?

Thanks for any input!


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