MessageAmp™ II-Bacteria RNA Amplification Kit

Catalog number: AM1790

Ambion™  Related applications: Microarray Analysis

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Description

The MessageAmp™ II-Bacteria RNA Amplification Kit is for amplification of bacterial RNA for highly sensitive, whole-genome expression analysis. The kit uses an in vitro transcription (IVT)–mediated linear amplification method optimized for use with bacterial RNA. Advantages of the MessageAmp™ II-Bacteria RNA Amplification Kit:

Maximum sensitivity—amplify as little as 10 ng of mRNA or 100 ng total RNA; a 10- to 500-fold reduction in the input RNA needed to probe microarrays
Highest yield of full-length transcripts—includes ArrayScript™ RT which produces up to twice the amount of full-length cDNA as M-MLV, AMV, or SuperScript® II
Stringently reliable data—reactions are highly reproducible, with Pearson's Correlation Coefficients >0.995

Using the MessageAmp™ II-Bacteria RNA Amplification Kit
This kit is the only commercially available kit for amplification of prokaryotic mRNA. The kit is compatible with any bacterial species and input RNA prepared from multiple sources, such as total RNA, bacterial mRNA or RNA. 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.

Accessory products
Bacterial mRNA can be enriched from total RNA using the MICROBExpress™ Bacterial mRNA Enrichment Kit (Cat. No. AM1905). Bacterial RNA can be separated from host cell mixtures with the MICROBEnrich™ Kit (Cat. No. AM1901).
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Product Size: 20 reactions
Includes Label or Dye: No
Reverse Transcriptase: ArrayScript™ Reverse Transcriptase
Sample Type (Specific): Total RNA (Bacterial), mRNA (Bacterial)

Contents & storage

Store the following at -20°C in a frost-free freezer:
• 22 µl PAP
• 22 µl 10X Poly(A) Tailing Buffer
• 12 µl Poly(A) Tailing ATP
• 22 µl T7 Oligo(dT) VN Primer
• 22 µl ArrayScript™ Reverse Transcriptase
• 22 µl RNase Inhibitor
• 22 µl 10X First Strand Buffer
• 170 µl dNTP Mix
• 210 µl 10X Second Strand Buffer
• 42 µl DNA Polymerase
• 22 µl RNase H
• 84 µl T7 Enzyme Mix
• 84 µl 10X T7 Reaction Buffer
• 84 µl each T7 ATP, CTP, GTP and UTP
• 1.75 ml Nuclease-free Water
• 10 µl Control RNA (1 mg/ml)

Store the following at room temperature:
• 30 ml Wash Buffer
• 7 ml cDNA Binding Buffer
• 9 ml aRNA Binding Buffer
• 20 aRNA Filter Cartridges
• 40 aRNA Collection Tubes
• 20 cDNA Filter Cartridges + Tubes
• 20 cDNA Elution Tubes • 10 ml Nuclease-free Water

Documents

Manuals & protocols

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