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SuperScript® II Reverse Transcriptase (RT) is an improved version of SuperScript® RT. It is a DNA polymerase that synthesizes a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. This enzyme is genetically engineered by the introduction of point mutations in the RNase H active center to reduce RNase H activity. This structural modification eliminates degradation of RNA molecules during first-strand cDNA synthesis and gives SuperScript® II RT superior performance characteristics. • Yield – Reduced RNase H activity results in greater first-strand cDNA yields • Size of cDNA – Generation of RT-PCR products up to 12 kb • Thermostability – Full activity at 42°C • Flexibility – Uses either total or poly(A)+ RNA
Performance and Quality Testing: SDS-PAGE purity; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product.
Unit Definition: One unit of SuperScript® II is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min. at 37°C using poly(A) • oligo(dT)12-18 as template • primer (3).
Unit Reaction Conditions: 50 mM Tris-HCl (pH 8.3), 40 mM KCl, 6 mM MgCl2, 1 mM DTT, 0.5 mM [3H]dTTP, 0.1 mM poly(A), 0.1 mM oligo(dT)12-18, 0.1 mg/ml BSA, and enzyme in 50 µl for 10 min. at 37°C.
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.
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