Stem cell tranfsfection - Electroporation or ca-phos precipitated DNA? (Apr/19/2006 )

Curious if anyone has experience with transfecting stem cells using either electroporation (the Amaxa Nucleofector or other equipment) or Ca-phos precipitated DNA?

Thanks,
Helena

---

Electroporation is my chosen method for getting DNA into ES cells. Works very well with constructs up to BAC size. I use an Equibio EasyjecT system set at 250V and 900uF or a double pulse that I cant quite remember offhand.

I have never tried Ca Phosphate and dont think I would bother!

Hope that helps.

---

Thanks for the advice, it's much appreciated

---

I can't get available the electroporation in our lab, can use lipo-based transfection?

---

I have tried lipofectamine 2000 for mouse ES, it seemed not so good. I did it according to the manufacturer's protocol and didn't do an optimization, maybe this is why i had so low efficiency.

We have a BTX ECM 2001 electroporation in our lab, but i cant find any protocol about mES transfection, even from its manufacturer . could anyone have experience give me some suggestions ?

any suggestions are welcome!

thanks in advance!

---

---

what about Fugene 6?

---

I´m workin with mouse ES cells and in my hands Fugene HD works super!!,
and i tried out many different transfection reagents.

---