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al RNA, ArrayScript often eliminates the need for two rounds of amplification, still generating sufficient cRNA for array analysis. This translates to significant savings of time and reagents.
Figure 2. ArrayScript Reverse Transcriptase Produces up to Twice the cRNA Yield 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 cRNA was purified by glass filter and eluted in nuclease-free water for analysis by Agilent 2100 bioanalyzer and NanoDrop Spectrophotometer.
ArrayScript is Suitable for all RT
Applications
Reverse
transcriptases are also used in RT-PCR, cDNA library
construction, and RLM-RACE. In most of these applications,
synthesis of full-length cDNA is a critical parameter that
affects final experimental results. For example, high yields
of full-length cDNA result in the generation of cDNA libraries
that more faithfully represent the cellular transcriptome. The
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