Extraction of gram positive rRNA without a centrifuge

[teresa_mogul at my-deja.com]

Hi all, appologies if I have already sent this

Does anyone know the best way to extract RNA from gram positive
bacteria. It is very important that I maintain the integrity of the RNA
as I'm looking for very good sensitivity values.The bacteria have been
grown in a medium that may contain RNases. I have been using a vacuum
manifold and guanidium thiocyanate for my gram negatives and this seems
to work well.

Questions:
Does lysozyme destroy RNA?
Does mercaptoethanol effect the activity of lysozyme?
Does heating and cooling destroy RNA?
What is a good anti-inhibitor agent to use in RT-PCR (BSA seems to be no
help?

Thaks for any help!

Regards Teresa

I


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