- Highly sensitive microarray analysis of bacterial mRNA
Description and Application:
The MessageAmp™ II-Bacteria Kit enables highly sensitive microarray analysis of bacterial mRNA with the least amount of starting material. This kit enables prokaryotic RNA amplification for whole genome expression analysis from bacterial samples. The kit employs an in vitro transcription (IVT)-mediated linear amplification method optimized for use with bacterial RNA.
Compared to conventional eukaryotic systems, the MessageAmp II-Bacteria Kit labeling requires orders-of-magnitude less input RNA and provides high reproducibility and dynamic range with standard labeling protocols such as direct labeling by reverse transcription or post-labeling reverse transcription (RT) reaction products. This kit is the only commercially available kit for amplification of prokaryotic mRNA.
This kit is compatible with any bacterial species and input RNA prepared from multiple sources, such as total RNA, bacterial mRNA enriched using Ambion’s MICROBExpress™ Kit, or bacterial RNA enriched from host cell mixtures with Ambion’s MICROBEnrich™ Kit.
MessageAmp™ II-Bacteria Kit Features:
- Maximum sensitivity. The MessageAmp II-Bacteria Kit amplifies as little as 10 ng of mRNA, or 100 ng total RNA, for microarray analysis. Less starting material directly translates into increased sensitivity. Ambion scientists have amplified RNA up to 15,000 times, with routine experiments representing a 10- to 500-fold reduction in the input RNA needed to probe microarrays.
- Highest yield of full-length transcripts.
First strand cDNA synthesis is powered by ArrayScript™ RT, which produces up to twice the amount of full-length cDNA when compared to M-MLV, AMV, or SuperScript™ II. Full length cDNA is the best way to ensure production of truly representative microarray samples.
- Stringently reliable data.
Comparison between sample reactions provides stringent reproducibility, with the Pearson’s Correlation Coefficients > 0.995. Reliability between experiments enables direct comparison and confidence of results.
aRNA generated by the MessageAmp II-Bacteria Kit can be labeled by either inclusion of a labeled nucleotide in the IVT reaction, or by reverse transcription of the aRNA in a reaction containing a labeled nucleotide. The MessageAmp II-Bacteria Kit can thus be used to prepare both 33P-labeled sense strand cDNA (for nylon PCR arrays) and biotin- or fluorescent dye-labeled aRNA for glass slide microarrays or Affymetrix GeneChip® arrays.
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