MessageAmp™ II-96 aRNA Amplification Kit is the first kit designed for high throughput amplification of mRNA and is based on the patented Van Gelder-Eberwine linear RNA amplification method. This kit takes advantage of extensive improvements incorporated into the MessageAmp™ II aRNA Kit combined with a new magnetic bead-based purification of both the cDNA and the aRNA.
The new microplate format available with MessageAmp II-96 provides a simple, streamlined procedure for the entire aRNA amplification process, including first and second strand cDNA synthesis, in vitro transcription, and aRNA labeling. Figure 1 illustrates aRNA yields obtained with either the manual or the fully-automated MessageAmp II-96 protocol at various HeLa total RNA input levels for both 4 and 14 hour IVT reactions. Figure 2 compares the correlation coefficients (% present calls on Affymetrix® Human Genome Focus Arrays) obtained using MessageAmp II-96 (Plate 1 and Plate 2) versus Ambion's standard single-tube MessageAmp II reaction.
Magnetic beads are used for both the cDNA and aRNA purification steps, eliminating the need for vacuum filtration or centrifugation. The use of beads also enables elution into smaller volumes, thus providing a higher concentration of aRNA. The entire process can be performed either manually or fully automated using a robot integrated with a thermal cycler and orbital shaker. Each kit contains sufficient reagents to amplify 100 samples, thus providing four samples for pre-testing prior to using a 96 well format.