HELP!! transformation problems

Patrick F.H. Lai pfhlai at LOOKSMART.COM
Fri Aug 7 20:48:54 EST 1998


---- Begin Original Message ----
Dear fellow scientists,

Since we don't have the means to perform electroporation and don't have=

the money to buy those ridiculous priced competent cells sold
commercially  I've to make them competent myself, but I've tried it
several times (different incubation times, fresh solutions etc) but
failed each time, so if someone out there can help me out I really woul=
d appreciate it.

Peter van Santbrink
Division of Biopharmaceutics
University of Leiden
The Netherlands
E-mail: santbrin at LACDR.leidenuniv.nl
--- End Original Message ----

Dear Dr. Peter,

I have been using the 'TransformAid Kit' from MBI Fermantas, 
<http://www.fermentas.com/>

I keep some untransformed jm109 in the fridge and passage the bugs
when I start my overnight ligation reactions.
The next day, i use the kit to 'encompetent' the JM109 (takes 30 to 45
minutes) and mix the bugs with a few microlitres of the ligation mix.
After incubating for a few minutes on ice, i spread the mixture on 
my agar plates.
I usually get more colonies than i need the following morning.
Few headaches, and it costs only Can$45 for 40 transformations here in
Toronto.
Is this cheap enough ?
BTW, don't use TOP10 cells, it rarely works with the kit in my hands.






Hope this is useful info.   :-)


Patrick F.H. Lai  < PFHLai at looksmart.com >
Graduate Student
University of Toronto
Toronto, Ontario, Canada





       
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