- Subject:Re: Quick transformation protocol wanted
-
- Date:Thu, 13 Feb 1997 18:22:47 0700
- From:crasmussen @ anat.med.ualberta.ca (Cohn Rasmussen) Organization:
University of Alberta
- Newsgroups:bionet.molbio.methds-reagnts
- In article , marcusj~helix.mgh.harvard.edu (Jeremy Marcus) wrote:
- >The transformation protocol portion of the procedure outlined
below is
- >well worth noting. As far as I know, it can be performed with
any
- >competent bugs (not just with cells prepared with CaCl2 a la
Pope Kent),
- >as long as the plates are warmed to 37 for 45 mm to 1 hr prior
to
- ~plating. While this admittedly doesn't save a _ton_ of time, the
main
- >advantage is that, by eliminating the heat shock step, you gain
an hour
- >that is free of obligations, so you can, for example, go to
a talk or go
- >eat dinner while the plate is warming, and then come back and
plate your
- >bugs.
- For quick transformation we take a fresh overnight of bacteria (200
ul) spin down, resuspend in 200 ul of TFB (newest Maniatis manual). Sit
15 mm in ice, add DNA, sit 30 mm in ice, heat shock 60 sec, plate directly
onto plates. Efficiency is crap but for shuttling plasmids it's great.