PC12 / puromycin

[Woong Choi]

Dear Netters:

I am trying to select PC12 cells which were transfected with pRT-On
recombinant. pRT-On has puromycin resistant marker. I did a kill-curve
experiment with various concentration of puromycin and found that even
high conc. of puromycin could not decrease the survival of PC12 cells no
less than 40%.

The manufacture's manual recommanded that 0.5-2 ug/ml of puromycin is
enough to select and maintain the stable transfectant. I am using 3
ug/ml of puromycin and control PC12 cells (not transfected) survived
this condition.

I want to hear those with similar experiences.
[1] Does PC12 cell line really resistant to puromycin?
[2] Is there any alternate chemicals for puromycin in this case?

Thank you in advance.

Woong Choi, M.D.
wchoi at med.chungbuk.ac.kr