lambda won't cut
cytogen at chmeds.ac.nz
Wed Sep 9 17:11:55 EST 1992
In article <9209091448.AA23264 at server.uga.edu>, mcgraw%gandal.dnet at SERVER.UGA.EDU writes:
> Hello netters,
> A colleague provided us with some lambda clone DNAs "purified" from plate
> lysates. They won't cut with EcoR1. We tried further purification with
> additional phenol/chloroform extractions. The uncut DNA looks good on a
> gel, and the OD's are good. The inserts should cut out with EcoR1 based on
> the way the library was made (R1 linkers). We haven't worked with lambda
> in quite a while, but I imagine that some of you out there are cutting
> yours successfully as you read this.
> What do you suggest?
> Mike Grosse
> McGraw lab
> University of Georgia
Have you tried adding spermidine to 4mM in your digetions? This can make a big
difference with lambda preps. Also, try cutting less DNA in a bigger volume.
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