yields of full-length cDNA compared to other reverse transcriptases (Figure 1). Figure 2 demonstrates that up to twice as much cRNA is synthesized when first-strand cDNA is generated with ArrayScript compared to wild type M-MLV, AMV, or other engineered M-MLV reverse transcriptases when low amounts of total RNA are used (~100 ng). This improved performance may eliminate the need for two rounds of cRNA amplification when working with small samples, thus reducing reagent cost and reaction time.

Second-strand cDNA Synthesis. The second-strand cDNA synthesis reaction has also been optimized specifically for the first-strand cDNA products generated with ArrayScript. This results in maximal conversion of first-strand cDNA into full-length double-stranded cDNA templates, thus increasing the average length of cRNA synthesized using MessageAmp II.

Figure 1. ArrayScript Reverse Transcriptase Synthesizes Intact cDNA Products as Long as 9 kb. cDNA synthesis was performed with 100 U of either ArrayScript RT, SuperScript II (SSII), SuperScript III (SSIII), or wild type (wt) M-MLV RT in a 20 l reaction volume containing a synthetic, poly(A)-tailed 9 kb transcript (1 g), 1X RT buffer, 0.5 M oligo(dT) primer, 0.5 mM dNTPs, 40 U RNase Inhibitor, and 0.1 l [α-³²P]dATP. Reactions were incubated for 1 hr at 42C and quenched with EDTA. The cDNA products were resolved on a 1% denaturing agarose gel and detected with a phosphorimager.

Figure 2. ArrayScript Reverse Transcriptase Produces up to Twice the aRNA Y
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