lambda won't cut

Martin Kennedy cytogen at
Wed Sep 9 17:11:55 EST 1992

In article <9209091448.AA23264 at>, 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

Mike - 

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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