RNA extraction of bacteria - (May/14/2007 )
I'm going to do RNA extraction of a soil gram negative bacteria, Agrobacterium rhizogenes.
Does anyone have a suitable protocol?
I really need any suggestion
I have been looking for the protocol online but couldnt find it
one of my supervisor mentioned that its very hard to extract RNA from bacteria and she didnt think it might work
I really hope to hear your opinion
Thanks in advance
Using a kit is way much easier. I know people using Trizol.
I used Qiagen kit. It has an additional solution to protect the RNA from degradation during lysing part.
Thanks a lot for your suggestion
I'll try that
ask some technical service from a kit to do that.
here is my exp :
once i aimed at extract RNA from cells.
Some bacterias contamination was there, but very few. I washed the cells 5 times and resulting under microscope was ok.
I extracted RNA from many samples including that one
The RNA was generally ok but degraded in this population.
So my guess is :
the RNA in cells wazs degraded because of their stress.
the bacterias wich have lot of RNases and proteases are hard to disrupt and so the RNAses are acting longer.
The point now :
you need to be sure your kit/trizol will lyzes efficiently and quickly the bacterias. So better is to ask to be sure your method is suitable.