cosmid prep info wanted

lsalati at wvnvms.wvnet.edu lsalati at wvnvms.wvnet.edu
Tue Sep 21 10:25:58 EST 1993


Hi,

I received a cosmid clone from another researcher and would like some
suggestions on methods used for amplification and preparation of more
DNA.  I have never worked with cosmids before so I thought I'd ask here.

The cosmid is pCOS2EMBL with an insert of about 50 kb.

1) Is there a preferred bacterial strain for growing cosmids or are
   they dependent on the vector involved?  We currently have DH5-alpha,
   JM109 and JM101 and SURE cells in the lab.

2) What method is used to transform bacteria with cosmid DNA?
   Electroporation?

3) How careful do you have to be when preparing DNA from the bacteria?
   Since the DNA is large I expect that I have to more careful to prevent
   breakage.  Are there any changes from a standard plasmid prep other
   than being careful in handling the lysate?

Any help is much appreciated.

Stephen Klautky



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