Isolating Bacterial RNA from Eukaryotic Host Cells
Challenges for Studying Bacterial Host Interactions
Start By Preserving RNA Expression Profiles
|Figure 2: Recommended Procedures for Purifying Bacterial mRNAs for Whole Genome Expression Analysis Following Bacteria-host Cell Interactions.|
Isolating Bacterial RNA From Host-Pathogen Mixtures
Following selective lysis of host cells, bacteria can be harvested and RNA purified with Ambion's RiboPure Bacteria Kit. RiboPure Bacteria is the only commercially available kit containing all reagents necessary for phenol-extraction and glass fiber filter purification of bacterial RNA. It is also compatible with RNAlater. RNA purified with RiboPure Bacteria is of extremely high quality and well suited for genome expression analysis with DNA arrays.
Enriching for Bacterial mRNA
The MICROBEnrich Kit employs a novel technology to selectively remove >90% of human, mouse, or rat total RNA from a mixed population of mammalian and prokaryotic RNA. The remaining bacterial RNA can then be used in downstream applications such as array analysis. The MICROBEnrich reaction is scalable (up to 100 µg). The kit can be used to remove a total of 500 µg of mammalian RNA from a mixed population of mammalian/bacterial RNA. Typically, the MICROBEnrich Kit is used for 20 reactions, each consisting of mixed RNA samples containing up to 25 µg of mouse, rat, or human RNA. For further enrichment of bacterial mRNA, 16S and 23S bacterial rRNA can be removed from the bacterial total RNA using Ambion's MICROBExpress™ Kit. The MICROBEnrich procedure can be seamlessly integrated with Ambion's MICROBExpress Kit for elimination of bacterial large subunit rRNAs by further enrichment of bacterial mRNA.
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