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Isolate High Quality Total RNA from Bacteria
As with yeast, bacterial gene expression patterns
can also be affected during enzymatic or chemical pretreatments used to
disrupt cell walls prior to RNA isolation. The RiboPure-Bacteria RNA Isolation
Kit uses no enzymatic pretreatments, allowing for faster processing, which
preserves RNA expression profiles. The kit utilizes the same type of protocol
combining an efficient glass bead/one step phenol mediated disruption
with the Vortex Adapter followed by a glass filter based RNA purification
step. The kit has been used at Ambion to isolate high quality RNA from
Bacillus subtilis, Staphylococcus aureus, Pseudomonas
aeruginosa, Escherichia coli, Campylobacter fetus, and
Rhodobacter sphaeroides and is compatible with most bacterial RNA
species (Figure 3). The RiboPure-Bacteria RNA Isolation Kit contains sufficient
reagents for 50 RNA isolations, each from 107 up to 109
bacteria.
Figure 3. High Quality Bacterial RNA Obtained with RiboPure-Bacteria Kit. Total RNA (2 g) was separated on a 1% agarose gel in formaldehyde/MOPS buffer. Samples were heated at 80C for 10 min in denaturing agarose gel loading buffer with 100 g/l EtBr before electrophoresis at 5 V/cm.
RiboPure-WBC: Scalable Protocol for Isolating High
Quality RNA from Blood
Isolating intact RNA from blood is challenging
because: 1) only white blood cells (WBCs) contain appreciable
amounts of RNA an
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