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Bacterial RNA Isolation From Infected Eukaryotic Hosts

e supernatant and can be precipitated with ethanol. Figure 1 shows an Agilent 2100 bioanalyzer electropherogram of a complex host-bacterial RNA population before and after bacterial RNA purification with MICROBEnrich. The RNA profile indicates nearly complete removal (>95%) of the host 18S and 28S rRNAs.

Figure 1. RNA Before and After Enrichment with MICROBEnrich Kit. Rat liver total RNA (25 g) and E. coli total RNA (2 g) were mixed together and subjected to the MICROBEnrich procedure. Another mixed sample of total RNA was subjected to a mock MICROBEnrich procedure in which the capture oligonucleotide mix was left out of the reaction. RNA was precipitated and an equal volume of each sample was separated on an RNA Nano LabChip with the Agilent 2100 bioanalyzer.


Removal of Bacterial rRNA
If desired, upon completion of the MICROBEnrich procedure bacterial mRNA can be purified from the total RNA through the seamless integration of Ambion's MICROBExpress technology. This technology enables >95% removal of prokaryotic 16S and 23S rRNAs (Figure 2). See the article "Purify mRNA From Bacteria!" for information on MICROBExpress.

Figure 2. RNA Before and After Enrichment with MICROBEnrich. Rat liver total RNA (25 g) and E. coli total RNA (2 g) were mixed together and subjected to the MICROBEnrich procedure with or without a further enrichment by MICROBExpress. The total RNA lane was subjected to a mock MICROBEnrich procedure in which the capture olig
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