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ArrayScript RT: Maximize Yields of Full-length cDNA


Get Full-length cDNA
Ambion scientists have engineered a mutant form of M-MLV reverse transcriptase to improve production of full-length cDNA. This engineered reverse transcriptase, ArrayScript (patent pending), produces significantly higher yields of full-length cDNA than wild type RTs, and rivals even the newest engineered RTs on the market (Figure 1). Efficient synthesis of full-length cDNA is important for microarray analysis; it results in low 3'/5' ratios, which are a hallmark of high quality microarray data.

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 [a-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.


Double the Yield in RNA Amplification
One of the most rigorous applications for an RT enzyme is RNA amplification from low amounts of total RNA. In this application, first-strand cDNA generated with ArrayScript resulted in up to twice the yield of amplified RNA compared to cDNA synthesized using wild type M-MLV, AMV, or other engineered M-MLV reverse transcriptases (Figure 2). When starting with low amounts of tot
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