Suitable for isolating bacterial RNA from mixed samples. The Ambion™ Kit includes sufficient reagents to remove 500 µg of host cell RNA from complex host bacterial RNA (20 reactions x 25 µg RNA). Analysis of gene expression following host-bacteria interactions is changing our understanding of host responses to bacterial infection. Many studies have used microarray analysis to study changes in the host cell transcriptome in interaction with bacteria. However, since there is no method to separate bacterial RNA from host cell RNA, corresponding analysis of bacterial transcriptomes is not possible. The MICROBEnrich™ Kit enables whole-genome microarray expression analysis of bacterial RNA following host-bacteria interactions.

Start with any total RNA sample
For microarray analysis and other sensitive applications, it is recommended before proceeding with the MICROBEnrich™ kit procedure DNase treatment to remove contaminating genomic DNA from total RNA.

Removes >90% of mammalian RNA from complex total RNA mixtures
MICROBEnrich™ Reagent The cartridgeuses hybrid capture technology (patent pending) to remove human, mouse and rat RNA (mRNA and rRNA) from complex host bacterial RNA populations, leaving behind enriched total microbial RNA. In the first step of the procedure, host bacterial total RNA is incubated with a mixture of capture oligonucleotides that bind mammalian 18S and 28S rRNA as well as polyadenylated RNA. Next, rRNA/oligo hybrids and all polyadenylated mRNA are removed from the mixture via oligonucleotide-derivatized magnetic beads. After magnetic separation, the remaining bacterial RNA in the supernatant is ethanol precipitated. Agilent 2100 Bioanalyzer electropherogram (shown) shows almost complete removal (>95%) of host 18S and 28S rRNA from the mixed sample. Bacterial mRNA purified using the MICROBEnrich™ kit is a better template than total RNA for downstream applications, so its sensitivity is significantly improved when used in applications such as array analysis (see figure) .

MICROBEnrich™ kit will work with any bacterial species
MICROBEnrich™ kit is optimized using E. coli RNA in the context of human, mouse and rat RNA. However, the kit should work well with RNA from any bacterial species. Although many mammalian host species are compatible with the MICROBEnrich™ Kit, only human, mouse, and rat rRNA sequences were used for capture oligonucleotide design and optimization.

Auxiliary products:
A magnetic stand is required to use this kit. They are available in a variety of sizes, including single-tube magnetic stands (SKU #AM10026), 6-tube magnetic stands (SKU #AM10055), and 96-well magnetic stands (SKU #AM10027 and #AM10050). RiboPure™ Bacteria Kit (SKU #AM1925) is suitable for purifying total RNA from host bacterial cells. TURBO™ DNA-free™ or DNA-free™ DNase Treatment and Removal Reagents (SKU #AM1907 and AM1906, respectively) are recommended for microarray analysis and other applications requiring contamination removal Applications of Genomic DNA. Following the MICROBEnrich™ procedure, bacterial mRNA can be purified from total RNA using the MICROBExpress™ Bacterial mRNA Enrichment Kit (SKU #AM1905). This technology removes >95% of prokaryotic 16S and 23S rRNA.