Help: PCR screen of colonies

KIM dkim at hector.NMSU.Edu
Wed Sep 15 20:43:15 EST 1999


RE:  Colony PCR:

It has been reported in this group that toothpicks may have an inhibitor
of the polymerase, and so plastic transfer implements are recommended.

In order to have an arrayed replica plate, I touch a colony with a yellow
tip, "spot" the tip three times on a plate which had been gridded for easy
reference, then place the tip into a PCR reaction mix of 10 microliters.
When a suitable number of colonies has been picked, "swizzle" the tips and
remove.  Add a tiny drop of oil (using a fine-bore plastic pipet) and put
in the machine.

Daniel Kim
dkim at nmsu.edu



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