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MICROBExpress (patent pending) is currently the
only commercially available bacterial mRNA purification kit. It uses a
novel strategy that rapidly removes >95% of the 16S and 23S rRNA from
total RNA of E. coli and other bacterial species
(Figure 1). MICROBExpress can be used with total
RNA obtained by a variety of methods, including glass fiber filter-based
methods (such as Ambion's RNAqueous Kits), and phenol-based methods (such
as Ambion's RiboPure-Bacteria Kit). The mRNA isolated with MICROBExpress
is a superior template for synthesizing labeled cDNA for array analysis
and is ideal for quantitative RT-PCR, Northern applications, and cDNA
library construction.
Quick and Specific Removal of rRNA
In the first step of the MICROBExpress procedure, total RNA is
mixed with an optimized set of capture oligonucleotides that bind to bacterial
16S and 23S rRNAs. Next, the rRNA hybrids are removed from the solution
using derivatized magnetic microbeads. The mRNA remains in the supernatant
and is recovered by ethanol precipitation. In addition to purifying bacterial
mRNA that is virtually free of 16S and 23S rRNA, this method also delivers
the benefits of quick processing (less than 2 hours), ease of handling,
and scalability.
Figure 1. Depletion of Bacterial 16S and 23S rRNA with MICROBExpress.
16S and 23S rRNAs were selectively removed from a 10 g E. coli
total RNA sample using the MICROBExpress Kit. A second 10 g sample
of the same total RNA prep was subjected to a m
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