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This reverse transcriptase or RNA-dependent DNA polymerase is used for real-time fluorescent RT-PCR with either SYBR Green dye or TaqMan probe.
The reverse transcriptase is a thermophilic type A polymerase (US patent pending) with optimal temperatures at 60-62°C (User Manual Tables 1-2).
The RTase is easily heat-inactivated at ≥90°C for 1min.
The enzyme efficiently synthesizes a complementary DNA strand on RNA template from a gene-specific primer, ≤1 unit per 20μL of singleplex reaction.
The enzyme reversely-transcribes single digit copies of target RNA molecules consistently.
The concentrations of the primers are variable depending on assay designs and thermocycling protocols (User Manual Table 4).
The preferable PCR product size is ≤150bp.
It is particularly suitable to one-step real-time qualitative and quantitative RT-PCR.
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