B cell transfections

[Melanie Brazil]

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


Melanie I. Brazil

Molecular Immunology Department 
National Institute for Medical Research
The Ridgeway
Mill Hill
London
NW7 1AA
UK

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

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