Summary: simple protocol for colony PCR

[Robert...@jcu.edu.au]

Thank you to those replying to my request for a simple protocol for carrying
out PCR on bacterial colonies. The essence of the replies was:

1. Either just add the cells that cling to a toothpick or tip to the PCR mix
or boil them prior to adding a small amount to the reaction mix.

From this combination I am going to try direct addition of cells (the initial
increase of temp to 95 dgc will be extended by 1 minute (----> heats the
cells ----> bust !)

2. A nice touch (pun intended) was to use the same tip or toothpick to touch an agar
plate after sloughing of a few cells in the PCR mix ----> new colonies in order
by the end of the day ready to pick for an overnight culture !!! (very quick)

3. The other factor that shone through was the need for extending the number of cycles
or by re-amplifying a portion of the first mix

I wont bore you with any actual PCR details

Hope it helps someone else too !!

Cheers,

Robert

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Dr Robert J. Coelen fax: 61-77-791 526
Dept of Biomedical and Tropical Veterinary Sciences phone: 61-77-815 024
James Cook University of North Queensland
Townsville-just-about-on-THE-reef, Q, 4811
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