( ield in MessageAmp II. A ...)RNA Amplification Just Got Better: MessageAmp II

ield in MessageAmp II. A comparison of ArrayScript RT (AS; 200 U), SuperScript II (SSII; 200 U); wild type M-MLV RT (200 U), and wild type AMV RT (10 U) in MessageAmp II conditions demonstrates that ArrayScript outperforms other commercial RTs. 90 ng HeLa-S3 total RNA (28S/18S ratio = 1.7) was heat denatured in the presence of T7 Oligo(dT) at 70C for 10 minutes. Reverse transcription was initiated with the RT enzyme and incubated at 42C for 2 hr in an air incubator. Following first strand cDNA synthesis, DNA polymerase was added for second strand synthesis, which was incubated at 16C for 2 hr in a thermocycler (heated lid option turned off). The double-stranded cDNA was then filter purified and added to a 4 hr in vitro transcription reaction. Last, the resulting aRNA was purified by glass filter and eluted in nuclease-free water for analysis by Agilent 2100 bioanalyzer and NanoDrop Spectrophotometer.

An Improved Amplification Kit
The RT and second-strand cDNA synthesis improvements, combined with the use of MEGAscript technology in the in vitro transcription reaction (IVT) result in the ability to use shortened IVT incubation times, and the generation of expression information from total RNA samples <1 g. In two round amplification reactions, the MessageAmp II Kit facilitates robust amplification of total RNA from samples as low as 100 pg.

The MessageAmp II aRNA Amplification Kit contains all the necessary reagents for first-strand cDNA synthesis, RNase H digestion, second-strand synthesis, cDNA purification, in vitr
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