B cell transfections

Melanie Brazil m-brazil at nimr.mrc.ac.uk
Mon Aug 7 06:26:57 EST 1995

Paul Yeo 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.

Dear Paul,
I transfect a B cell line using protoplast fusion. It is basically the
protocol from Maniatis. I found that my b cell line (LK35) needs a high
concentration for selection - I use 2mg/ml. I find that only 50% of my
transfections work - ie I get colonies, and then the efficiency is low
(5-10 colonies / 5x106 cells)
Good luck 

Melanie I. Brazil

Molecular Immunology Department 
National Institute for Medical Research
The Ridgeway
Mill Hill

tel: (44 181) 959 3666 x2457
fax: (44 181) 913 8531

email: m-brazil at nimr.mrc.ac.uk

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