Perform Microarray Analysis with Limited Bacterial RNA
Over the past decade, analysis of whole-genome expression profiling with DNA microarrays has revolutionized the way biologists study gene function. RNA amplification has become the preferred method of preparing probes for most microarray platforms. Unfortunately, the benefits of RNA amplification have not been easily accessible to researchers analyzing bacterial gene expression profiles. The absence of poly(A) tails on bacterial mRNA prevents the use of oligo(dT) primed reverse transcription, which is at the heart of most amplification methods. The lack of an appropriate method for bacterial RNA amplification has hampered studies that rely on rare or limited samples, such as environmental collections, small volume cultures, and bacterial RNA isolated from host cells. With over 160 microbial genomes fully sequenced, and many more in progress, an unprecedented opportunity to perform global expression analysis on important pathogens and model organisms is now available.
MessageAmp™ II-Bacteria Kit
Linear Bacterial RNA Amplification
Figure 1. Schematic of MessageAmp™ II-Bacteria Procedure.
Yields of Amplified RNA
Whole Genome Microarray Analysis
Tips for Bacterial RNA Amplification
A Complete Kit for Bacterial RNA Amplification
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 .