Schneider cell transfection

Emmanuel Skoufos, Ph.D. skoufos at nefeli.imbb.forth.gr
Mon Apr 24 21:36:13 EST 1995


Cam Patterson writes:
>I am planning to start transfections in Schneider cells
>to test transactivation of a reporter construct by Sp1 
>and AP2. Our lab has experience transfecting a variety of
>cell types, however no one has any experience with these
>cells. I have noticed that calcium phosphate and lipofection
>transfection techniques have been used with the Schneider
>cells. I was wondering if anyone out there who has experience
>with these cells could offer suggestions about the most
>efficient method for transfecting these cells, and any
>pitfalls, if any, that I am likely to encounter.
>Thanks in advance.
>
>cam

Hi,

We have recently started working with transfections of Schneider cells.  We
tried both electroporation and CaPO4 precipitation and did several test runs
calculating transfection efficiency using a Lac-Z expressing vector as control.
Our conclusions were 1.CaPO4 gave about a 5-10x better tranfectability (we did
not try lipofection) than electroporation 2.Qiagen preps were fine to use,
whereas "regular" midi-prep and miniprep DNAs didn't work well 3.It is
important for the cells to look "healthy" for the transfections to work 4.
Some media work better to keep cells happy (We had our best luck with media
made by Sigma especially for these cells).
Hope this helps,

Sincerely,

Emmanuel



  
_______________________________________________________________________________
Emmanuel Skoufos, Ph.D.                          | The opinions and views
Insect Molecular Biology Group                   | presented here are my own
Institute of Molecular Biology and Biotechnology | and only my own.
Box 1527                                         | --------------------------
Heraklion, Crete 71110                           |skoufos at nefeli.imbb.forth.gr
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Phone xx-3081-212890       FAX: xx-3081-231308   |ga119 at kanga.ins.cwru.edu



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