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Simplified, Efficient, Single-Round RNA Amplification for Microarray Analysis
The Ambion® MessageAmp™ Premier RNA Amplification Kit (magnetic bead RNA clean-up) is the latest innovation for the preparation of RNA samples for microarray analysis. This kit builds upon the linear RNA amplification method and reagents developed for the MessageAmp II Kits with enhancements that create:
NEW MessageAmp™ Premier RNA Amplification Kit
NEW MessageAmp™ Premier RNA Amplification Kit
- A simplified workflow (e.g., simplified protocol; streamlined reagents; single-tube format)
- Improved performance (e.g., up to 50,000-fold amplification using T7 MEGAscript® in vitro transcription technology)
- More flexible RNA input requirements (e.g., 20–500 ng total RNA)
All MessageAmp kits employ the proven methodology for RNA sample preparation and labeling based on the original T7 in vitro transcription (IVT) amplification technology, also known as the Eberwine or reverse transcription-IVT (RT-IVT) method [1].
The new Ambion MessageAmp Premier RNA Amplification Kit features magnetic bead-based RNA purification to yield sufficient biotinylated antisense mRNA for Affymetrix® GeneChips® from just nanogram inputs of total RNA (Figure 1). The kit incorporates extensive improvements to both workflow and sensitivity. A series of master mixes reduces the number of tubes, pipetting steps, and overall protocol steps. The result is a kit that produces enough amplified RNA (aRNA) for microarray analysis from as little as 20–25 ng of input RNA. The entire procedure from RNA amplification through hybridization to a GeneChip Array can be completed in a single day.
Figure 1. MessageAmp™ Premier RNA Amplification Kit Enables Researcher to Generate Sufficient aRNA for GeneChip® Array Hybridization Starting with Nanograms of Total RNA. Triplicate RNA samples (20 or 100 ng) from 7 tissues (Ambion® FirstChoice® Total RNA) or HeLa cells were amplified and labeled with the MessageAmp Premier Kit. Shown are the average amplified RNA yields (bars) and median size (diamonds) after 4, 8, or 14 hour in vitro transcription reactions.
This new simplified workflow of the MessageAmp Premier Kit significantly reduces hands-on time while offering improved sensitivity at an affordable price. Once purified, the aRNA is suitable for use on microarray gene expression systems designed for biotin-modified antisense RNA samples.
The new Ambion MessageAmp Premier RNA Amplification Kit features magnetic bead-based RNA purification to yield sufficient biotinylated antisense mRNA for Affymetrix® GeneChips® from just nanogram inputs of total RNA (Figure 1). The kit incorporates extensive improvements to both workflow and sensitivity. A series of master mixes reduces the number of tubes, pipetting steps, and overall protocol steps. The result is a kit that produces enough amplified RNA (aRNA) for microarray analysis from as little as 20–25 ng of input RNA. The entire procedure from RNA amplification through hybridization to a GeneChip Array can be completed in a single day.
References
- Van Gelder RN, von Xastrow ME, Yool A, Dement DC, Barchas JD, Eberwine JH (1990) Amplified RNA synthesized from limited quantities of heterogeneous cDNA. Proc Natl Acad Sci USA 87:1663–1667.