Water or TSB to dilute an over night culture!? - (Dec/27/2005 )

When diluting an over night culture of bacteria for the PCR template, should water or TSB (media) be used?

I usually dilute it in TE

Spin down the cells and resuspend in sterile water. This will get rid of extracellular enzymes, some of which may be nucleases. I spin down 1 ml of cells and then resuspend in 200 ul of dH2O.

depends on the bacteria...for S aureus, we use TEN to resuspend the cells (then there is a hairy procedure to lyse the cells, then they are boiled, then 2 ul goes to PCR)...if you just resuspend the cells and stick them into a PCR reaction, you won't see anything

so, I think it depends on the cells you are working with.

depends on the bacteria...for S aureus, we use TEN to resuspend the cells (then there is a hairy procedure to lyse the cells, then they are boiled, then 2 ul goes to PCR)...if you just resuspend the cells and stick them into a PCR reaction, you won't see anything

so, I think it depends on the cells you are working with.

It depends on the bacteria. Plenty of bacteria survive fresh water...they may be in osmotic shock but they are not necessarily dead. And besides, does it really matter so long as the DNA is intact? That's all you need for the PCR. If you are unsure, try both methods...resuspend one set of cells in TSB and another in water and do a PCR, it will take half a day.