Reliable RT for Full-length cDNA Synthesis and High Yields

Description:
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 rather than a deletion in the RNase H active center. Like SuperScript™ RT, SuperScript™ II RT has reduced RNase H activity. Unlike SuperScript™ RT, however, the selective mutations within the RNase H domain maintain full polymerase activity. This structural modification eliminates degradation of RNA molecules during first-strand cDNA synthesis and gives SuperScript™ II RT superior performance characteristics, including:

• Greater first-strand cDNA yields
• More full-length cDNA synthesis
• Full activity at 42°C

Source: Purified from E. coli expressing the pol gene of M-MLV (1,2), mutagenized to reduce the RNase H activity.

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 eoxyribonucleotide into acid-precipitable material in 10 min. at 37°C using poly(A)•oligo(dT)12-18 as template•primer.

Unit Reaction Conditions: 50 mM Tris-HC1 (pH 8.3), 40 mM KC1, 6 mM MgC1 2 , 1 mM DTT, 0.5 mM [3H]dTTP, 0.1 mM poly(A), 0.1 mM oligo(dT)12-18 , 0.1 mg/m1 BSA, and enzyme in 50 µl for 10 min. at 37°C.

Contents and Storage:
SuperScript II is supplied with a vial of 5X first-strand buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2 ], and a vial of 100 mM DTT. Store at -20°C. Guaranteed stable for 6 months when properly stored.