Transfection of B cell-lines

Mike Smith smith-m at oberon.hsc.colorado.edu
Tue Aug 8 10:51:12 EST 1995


Laurentiu COCEA <cocea at citi2.fr> wrote:
>r.p.yeo at reading.ac.uk wrote:
>>
>>    I am trying to transfect Daudi and Raji cell-lines with a number of
>> expression vectors with either HCMV or RSV LTR promotors with very little
>> success.  Is there anybody out there with experience in transfecting these
>> lines either transiently or stable and could they email me a procotol. 
>> Thanking you in advance.
>> 
>> Paul Yeo
>> AMS,
>> The University of Reading
>> Whiteknights, PO BOX 228
>> READING, RG6 6AJ

I've had good success with Raji (haven't tried Daudi) using 
electroporation.  Using the BioRad or BTX machines.  
500V/220 ufd capacitance in a 4 mm cuvette.  Used 20ug DNA 
in 200ul of cells in RPMI1640+10%FBS.  If you want any more 
details email me.  Good Luck

Mike Smith, Ph.D.
Univ. of Colorado HSC




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