Buffer for plaque phage lysis --> PCR ?

[Stuart Brown]

In article <37h8tc$kb7 at guru.med.cornell.edu>
fgrun at med.cornell.edu (fgrun) writes:
 
>Does anyone have a buffer recipe that allows you to pick and lyse
>single lambda-gt10 plaques for PCR screening ?
>Would TE pH 8.0 + 1% SDS at 70 C for 5 mins be sufficient to remove the
>phage coat and allow efficient priming ?
>Thanks.
>
>Felix Grun
>fgrun at mail.med.cornell.edu
 
 
I have had no trouble at all doing PCR on phage plaques picked with a
Pasteur pipet (poke through the top agarose into the agar below and lift
out a tiny plug from the center of each plaque).  I blow the plug of agarose
out into 500 ul of SM buffer (Tris, salt and gelatin) allow phage to diffuse
out for an hour or so, then take 1 ul as template for PCR reaction.  I
make no changes to my PCR buffers or cycle conditionss as compared to
purified plasmid template.
 
Stuart Brown                       |   Plant Genetic Resources
                                   |   Georgia Experiment Station
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