Help on transfecting JURKAT!

[Javier Quintero Vazquez]

One collegue of my lab request to me these question:

Please send the mail to:
	Dr. Primitivo Barja
	Email: bnprimi at usc.es

We are electroporating JURKART cells to introduce an episomal expresion 
vector which contains the hygromicin resistence gene as selection method.
We have had several problems and we are looking for some help.

There is anyone information about:

1.- Electroporation conditions about JURKAT cells.
2.- Growth conditions of transfected JURKAT in medium containing hygromicin
(range of hygromycin concentrations at which JURKAT cells survive or are 
killed).
3.- Detection and quantification of the transfected plasmid (We need, if 
possible, a single protocol to detect it, for example by PCR).
4.- How to determine the frequency of tranfected cells.

Thanks, in advance.
  
Dpt. Biochemistry & Molecular Biology. 
University of Santiago de Compostela
SPAIN