My lab is using PCR to detect bacteria in environment samples, however, PCR cannot distinguish between dead or live bacteria. I found that some papers reported that ethidium monoazide could help, but it seems no use in my experiment. Shall I use PMA instead of EMA for it?
My experiment setup is like this:
1) 1ml of bacterial culture was heat treated at 72c for 15mins (confirmed no growth in agar)
2) EMA (100mg/ml) added to culture
3) expose to 650W halogen light for 1 min
4) pellet the cell at 5000g x 5mins
5) extract DNA
6) PCR
However, still got a very bright band even I diluted culture to 0.1McF......