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Bacterial Whole Genome Array Analysis

elds of exceptionally pure RNA.


Removing Contaminating Genomic DNA: TURBO DNA-free
Once the RNA is purified, it should be treated with DNase to remove any contaminating genomic DNA. This is especially important for array analysis and/or qRT-PCR. Contaminating genomic DNA will be labeled during the cDNA synthesis/labeling reaction and/or lead to non-specific RT-PCR products that could interfere with quantitative results. TURBO DNA-free is ideal for this application (see The World's Best DNase).


Enriching for Bacterial mRNA: MICROBExpress
For decades mRNA has been isolated from eukaryotic sources using oligo(dT) selection. Bacterial mRNAs, however, lack the relatively stable poly(A) tails found on eukaryotic messages and thus cannot be enriched using affinity selection with oligo(dT). Ambion's MICROBExpress Kit offers enhanced signal sensitivity in gene expression analysis across all array platforms tested (including glass, nylon and GeneChip) by its ability to remove >95% of bacterial rRNA from total bacterial RNA preparations. The depletion of rRNA leaves behind a highly enriched bacterial mRNA population (See Figures 1 and Figure 2). Additionally, MICROBExpress leads to an increase in the total number of genes detected (i.e. increase in % present calls) and an improvement in overall signal to noise on GeneChip E. coli Antisense Genome Arrays (Figure 3).


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