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Transformation problems...

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Mon Jan 17 12:24:58 EST 2000


Jan-Henner Wurmbach <wurmbach at uke.uni-hamburg.de> wrote:
> "R. Jayakumar" <jakku at mrna.tn.nic.in> schrieb im Beitrag
> <004601bf6031$e485c820$8c01a8c0 at tn.nic.in>...
> [snip]
> 
>>     ONE STEP TRANSFORMATION PROTOCOL FOR E.COLI
> [snip ]
>> 5. Take a 100ul aliquot and add DNA (around 10-20ng) and leave on ice for
>> 30min (we found that heat shock treatment gives less transformants than
>> ice treatment).
> Interesting! In our lab Heat shock is needed, at least with ligations.

If you use the Chuang protocol (which the previous poster did), heat
shocking is detrimental for transfection efficiency in my experience
as well. If you use some Inoye (sp?) variant, heat shock, on the other
hand, is essential for getting good transformation efficiencies, as
far as I can remember (haven't used it since a long time).

--Cornelius.

-- 
/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
/* "Science is the game we play with God to find out what His rules are."  */




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