about transformation of DNA into E.Coli

Paul N Hengen pnh at ncifcrf.gov
Wed Aug 7 15:21:19 EST 1996


Sailesh Surapureddi (Sailesh.Surapureddi at mcb.liu.se) wrote:

> 1. Pre warm the plates @37 (very imp) for 30min or 60min which ever is
> convient to you.
> ...
> 9. Take the entire amount and spread  on the prewarmed LB +antibiotic
> plates  (no SOC etc etc).
 
In my opinion, you are still heat shocking your cells by plating on
pre-warmed media. I've seen people heat shock at 37C for 10 min. before
plating and they swear it works better than what I normally do, which
is 42C for 90 sec. At one time I did a side-by-side comparison and
didn't see any difference, so I never switched. I'd really like to see
your numbers of transformants per ug DNA compared with other times
and temperature of heat shock if you have any.

--
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