Large-ish vectors that won't behave

Sasha Kraev bckraev at aeolus.ethz.ch
Wed Nov 23 06:51:06 EST 1994


I had exactly the same problem several years ago, when I needed to routinely
transfer a 2 kb cassette into a 12 kb vector. I overcame it by transforming XL-1
or AG-1 ( both Stratagene), which I made competent acc. to Hanahan and pre-
tested with pTZ19 ( about the same as pGEM) to have efficiency of >2.10E7.
I transformed 50 ng of ligated DNA and plated on two 90 mm plates, which gave
5-20 colonies. From those I always had at least one correct clone. Hope this
helps. Sasha



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