Quick transformation protocol wanted
Paul N Hengen
pnh at ncifcrf.gov
Tue Feb 18 16:36:59 EST 1997
A.P.N. de Boer (deboer at nefeli.imbb.forth.gr) wrote:
> The fastest I know of is the method of Chung which uses a solution of
> PEG and DMSO made in LB. For very efficient transformation you grow
> cells to mid log, chill on ice, centrifuge and resuspend in 1/10 vol of
> solution. They are ready to use or to freeze. For quick and dirties you
> can make up the solution as a 2X stock and just add an equal volume to
> your cells, no need to centrifuge or anything. It even works with
> overnight cultures, although at reduced efficiencies.
How about resuspending a colony in 1X TSS? Anyone do it this way?
Wasn't there a protocol for resuspending a colony and doing a quick
freeze - thaw, add DNA and plate? I think that's the one you're looking for.
--
*******************************************************************************
* Paul N. Hengen, Ph.D. /--------------------------/*
* National Cancer Institute |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology | Phone: (301) 846-5581 |*
* Frederick Cancer Research and Development Center| FAX: (301) 846-5598 |*
* Frederick, Maryland 21702-1201 USA /--------------------------/*
* - - - Methods FAQ list -> ftp://ftp.ncifcrf.gov/pub/methods/FAQlist - - - *
* - TIBS column archive -> http://www-lmmb.ncifcrf.gov/~pnh/readme.html - - *
* - The BEST Molecular Biology HomePage -> http://www-lmmb.ncifcrf.gov/~pnh/ *
*******************************************************************************
More information about the Methods
mailing list