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Bacterial RNA Isolation From Infected Eukaryotic Hosts

ure. This host-bacterial cell total RNA mixture can be obtained by a variety of RNA isolation methods, ideally by Ambion's new RiboPure-Bacteria Kit. Other methods including glass fiber filter based methods (such as Ambion's RNAqueous Kits) and phenol based methods (such as Ambion's RNAwiz and the ToTALLY RNA Kit) are also appropriate. For microarray analysis and other sensitive applications, Ambion also recommends a DNase treatment (such as with DNAfree Treatment and Removal Reagents) to remove contaminating genomic DNA from the total RNA prior to the MICROBEnrich procedure.


Remove >90% of Mammalian RNA from Complex Total RNA Mixtures

MICROBEnrich employs hybridization capture technology (patent pending) to remove human, mouse, and rat RNA (both mRNA and rRNA) from complex host-bacterial RNA populations, leaving behind enriched microbial total RNA. In the first step of the MICROBEnrich procedure, host-bacterial total RNA is incubated with an optimized mixture of capture oligonucleotides that bind to the mammalian 18S and 28S rRNAs and polyadenylated RNAs. Next, the rRNA/oligo nucleotide hybrids and all polyadenylated mRNAs are removed from the mixture with oligonucleotide-derivatized magnetic beads. After magnetic separation, the bacterial RNA remaining in th
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