TRANSFER OF PLASMIDS INTO BACTERIA

Kevin Oldenburg kevin_oldenburg at qmgates.affymax.com
Fri Mar 3 11:44:42 EST 1995


You will want to transfect a bacterial strain with each of the plasmids,
and then freeze down the bacteria in 50% glycerol in order to maintain a
permanent stock.  I would first add approximately 20ul of sterile dH2O to
each of the samples in order to increase the volume.  If you have access
to an electroporator go ahead and electroporate 1ul of the plasmid stock
into 40ul of competent bacteria (call Bio Rad for a protocol if you don't
already have one).  If you don't have an electroporator then go with the
Calcium chloride method (ref. Molecular Cloning: A Laboratory Manual  Au:
Maniatis Pg:1.82). I highly recommend this molecular biology manual.  Good
Luck
Kevin Oldenburg

In article <9503021338.aa06191 at etsuodt.etsu.edu>, betts at ORION.ETSU.EDU
(GORDON BETTS) wrote:

> I have obtained a pair of plasmids (a whole 1 ul of each) and now must do
> something with them.  Since I am new to this area of work, I'm hesitant to
> just jump in.  So, does anyone have good advice on how to do a one-time,
> not-a-chance-of-failure transfection into bacteria?  What's a good vector
> for this?
> Thanks,
> Gordon Betts
> Biology Dept.
> East Texas State University
> Commerce, TX 75429-3011
> 903/886-5369
> ============================================================================
> ======
> Life is like a bowl of stew, you need to stir things up once in a while so 
> the scum doesn't rise to the top.
>       Author unknown
> ============================================================================
> ======

-- 
The views expressed here are my own and are not necessarily those of my company.



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