Extract DNA from dead bacteria - (Dec/08/2006 )
Hi all,
I have a bacteria strain containing my plasmid spread on LB plate four months ago.
I forgot to make freezing stock of the bacteria, can anybody tell me how to get my plasmid out from the bacteria??
Thank you !
Elaine
Before you assume they are dead, try to regrow them (with antibiotic).
Bacteria are pretty resilient.
If you are unable to grow the bacteria, you can always do a quick transformation with the DNA and plate a new batch. If you do not have the DNA, you can try doing a plasmid prep from a single colony. However, I wouldn't trust this DNA since the cells have been under prolonged stress and also your antibiotic is going to be at least partially inactive. Good luck!
I have a bacteria strain containing my plasmid spread on LB plate four months ago.
I forgot to make freezing stock of the bacteria, can anybody tell me how to get my plasmid out from the bacteria??
Thank you !
Elaine
I would try doing a prep and/or just resuspending some colonies in H20 and heating and freezing, then transform in high competent cells.
If E.coli, must probably b dead by now.
Pick a colony, do a miniprep and transform.
though it seems a minute amount of bacteria left, remember all it takes is 1 surviving plasmid. Keep trying if its important, or else order some new stock.