Ca-PO4-transfection

Wolfgang Schechinger root at 127.0.0.1
Sat May 11 00:17:02 EST 2002


Dear Alexandra, 

You might check the Nolan's Lab Retroviral Pages for details on CaPO4:
http://www.stanford.edu/group/nolan/protocols/pro_helper_dep.html

or the simpler procedure in in this paper (it's from my former colleagues and 
I know it works very well for 293 HEK cells. If you have difficulties to 
locate a copy, please mail me)

http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_ui
ds=11781100&dopt=Abstract

The crucial thing is to get a *fine* precipitate that is almost invisible 
under the microscope. The first I did it, I thought I had tons of E. coli 
growing.

If you're looking for another cheap procedure (one bathtube of reagent is 50 
EUR, consider the transfection of whole cows :), you might try the 
polyethyleneimine (PEI) protocols sometimes discussed in this NG. They work 
great for me on 3T3 cells when e.g. screening large numbers of minipreps, when 
speed and costs are more important than efficiency. Check the NG's archieves 
at http://www.bio.net/hypermail/METHDS-REAGNTS/

Regards and good luck,
Wo

hubahopp at gmx dot de

> Hallo everybody,
> 
> can soebody help in kind of CaPO4-transfection und tell me some details
> about buffers, amounts of DNA, CaPO4 and buffers for an optimal
> transfection result?
> 
> 


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Dr. Wolfgang Schechinger
Institute of Biomedical Sciences
Academia Sinica, Taipei, Taiwan
E-mail wolfsc at ibms dot sinica dot edu dot tw

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