mRNA extraction from Gram positive bacteria
graham at biodec.wustl.edu
Sun Jun 2 12:45:51 EST 1996
I think you will find a oscillating or high-speed reciprocating
shaker almost essential for efficient isolation from many gram
positives. These devices shake the pelleted suspention at high
speeds (6K rpm) with zirconimum fragments (beads) and will increase
the total RNA yield from say Mycobacterium almost 10X compared to
other grinding and lysis procedures, in my experience. Two such
devices are "Fast-prep" from Gibco and "Mini Bead Beater".
As for Trizol, its just colored buffered phenol of low PH, with a
buffer system that supposedly allows recovery of the DNA from the
organic phase. Published reports show that up to 20% of the nucleic
acids recovered from the aqueous phase may be DNA.
See Fischetti's recent Analytical Biochemisty article for details.
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