01 January 2013 35 8K Report

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......

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