Bacteria Have mRNA Too

Until recently, scientists were largely limited in their choice of mRNA sources. For decades it was possible to isolate mRNA from eukaryotic samples, such as animal and plant cells, but it was virtually impossible to isolate bacterial mRNA. That's because bacterial transcripts lack the poly-(A) tails found on eukaryotic messages, upon which oligo-(dT) selection, the traditional eukaryotic purification technique, is based. But now researchers studying bacterial RNA have a new option. Austin, Tex

Deborah Stull

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But now researchers studying bacterial RNA have a new option. Austin, Texas-based Ambion Inc.'s MICROBExpress™ Bacterial mRNA Purification Kit, offers researchers a method for isolating mRNA from many bacterial species, including both Gram-negative and Gram-positive organisms, with greater than 95% of the 16S and 23S rRNA removed.

Unlike the positive, oligo-(dT)-based techniques used to purify eukaryotic transcripts, the MICROBExpress protocol is a negative selection technique, enriching for bacterial mRNA by removing the larger rRNA species that comprise 80% or more of total RNA samples. Researchers start by isolating total RNA from the bacteria of interest using any of a variety of techniques, including glass fiber filter- or phenol-based methods. This total RNA is then mixed with a set of capture oligos that have been optimized for binding to the bacterial 16S and 23S rRNAs. These rRNA hybrids are removed using derivatized magnetic beads, leaving the mRNA in the solution to ...

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