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Perform Microarray Analysis with Limited Bacterial RNA

not be compared directly with amplified samples. All amplification methods, or any labeling method for that matter, produce some biases in mRNA representation. What is important for expression analysis is that the biases are reproducible within a method and that similar labeling methods are used on samples being compared. MessageAmp II-Bacteria compares favorably with standard labeling protocols in terms of reproducibility and dynamic range, but with a dramatically reduced requirement for input RNA.


A Complete Kit for Bacterial RNA Amplification
The MessageAmp II-Bacteria Kit provides reagents for 20 RNA amplifications starting with as little as 10 ng bacterial mRNA or as much as 500 ng total RNA. A variety of labeling methods can be used with the kit, including 33P cDNA labeling, direct labeling with Cy dyes or biotin, and indirect coupling using aminoallyl nucleotides (modified nucleotides are not provided with the kit). E. coli total RNA is provided as a control.

While MessageAmp II-Bacteria can be successfully used with total RNA, MICROBExpress enriched bacterial mRNA is a much better choice for microarray analysis. A 10-fold to 100-fold increase in sensitivity is seen when using enriched mRNA, compared with total RNA. Overall signal and signal-to-noise ratios can be dramatically increased by using RNA prepared using the MICROBEnrich and/or MICROBExpress Kits with MessageAmp II-Bacteria [1].
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