Electroporation Challenge!!!

Antonio Rodriguez Franco bb1rofra at seneca.uco.es
Mon Jul 17 12:36:06 EST 1995


In article <3u1ktn$l3v at sifon.cc.mcgill.ca> Graham Dellaire <popa0206 at PO-Box.McGill.CA> writes:
>From: Graham Dellaire <popa0206 at PO-Box.McGill.CA>
>Subject: Electroporation Challenge!!!
>Date: 12 Jul 1995 23:12:55 GMT

>HAs anyone out there obtained an electroporation frequency
>better than 10-5 transformants/no# cells transformed???

>If you have I would like to find out if your cell type and
>what make of Gene Pulser you have etc.... 

>I am having a hell of a time breaking the 10-5 barrier.


>Graham


>_______________________________________________________________________ 
>Graham Dellaire                     Snail Mail:
>                                    Red Cross, Research         
>McGill University                   Montreal Blood Services             
>Faculty of Medicine                 3131 Sherbrooke St. East         
>Div. of Experimental Medicine       Montreal, QC, Canada           
>E-mail: popa0206 at po-box.mcgill.ca   H1W 1B2                        
>B2XE at musicb.mcgill.ca                                                      
>WWW Page: http://www.medcor.mcgill.ca/EXPMED/expmed.html        
>Fax: (514) 525 0881                                                             
>Voice: (514) 527 1501 ext 175                                             
>_______________________________________________________________________

We have increase the eficiency of electroporation about 1000 times with the 
following tricks.

i) 2YT is better than LB medium to grow the cells
ii) After growing the cells, (100-250 ml) leave them in ice for around 1-2 
hours.
iii) do not use water in your centrifuging and washes. Use water plus 10 % 
glicerol instead for every single step.

It seems that bacteria (young bacteria) cells are disrupted very easily, and 
do not survive to the treatment.

Hoping this help,



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