TRISOLV RNA extraction from Gram +ves

mic268a at vaxc.cc.monash.edu.au mic268a at vaxc.cc.monash.edu.au
Tue Jan 25 20:32:50 EST 1994



Hi there,

Just wondering if anyone can help me with isolation of RNA especially
from Gram positives such as Clostridium spp.

I am using the TRISOLV method and of a number of different attempts
I have only one successful RNA prep. The RNA I get is usually degraded
or I get very low yields. 

Does anyone have any tips or suggestions?
I am growing the cells to an OD 580nm of 0.5 and 1.0 and am finding no
difference between the yields ie, they're both crap! Is the less cells
that you use the better or do you need more cells? Is OD important?
Can you pretreat the cells with lysozyme or something similar?

Any comments will be gratefully received

Thank You
Mike L
Monash Uni
Australia






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