SuperScript® II Reverse Transcriptase

Catalog number: 18064022

Invitrogen™  Related applications: Reverse Transcription

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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 from 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 RT 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.

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 for use in diagnostic procedures.

Specifications

Optimal Reaction Temperature: 42° C
Final Product Size(s): 12.3 kb or less
Reverse Transcriptase: SuperScript® II
Ribonuclease H Activity: Reduced
Form: Frozen
Format: Tube(s)
Quantity: 2000 units
Template: ssDNA, ssRNA, RNA-DNA Hybrid
Sensitivity: Medium
Product Size: 2,000 units
Concentration: 200 U⁄µl
Enzyme Function: RNA dependent DNA Polymerase
Shipping Condition: Wet Ice
Number of Reactions: 10 Reactions

Contents & storage

SuperScript® II Reverse Transcriptase (2,000 units total, at 200U/µL) 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.

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