SuperScript® III Reverse Transcriptase

Catalog number: 18080093

Invitrogen™  Related applications: Reverse Transcription

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Description

SuperScript® III Reverse Transcriptase (RT) is a proprietary mutant of SuperScript® II RT that is active at 50°C and has a half-life of 220 minutes, providing increased specificity with gene-specific primers (GSPs) and the highest cDNA yield of all RTs. It is ideal for RT-PCR of a specific gene or for generating cDNA from total or poly (A)+ RNA samples. Like SuperScript® II, it synthesizes a complementary DNA strand from single-stranded RNA, DNA, or from an RNA:DNA hybrid. SuperScript® III RT is genetically engineered by the introduction of point mutations that increase half-life, reduce RNase activity, and increase thermal stability. SuperScript® III RT features:

Thermostability—half-life of 220 min at 50°C
Yield—reduced RNase H activity for more full-length cDNA
Specificity—full activity at 50°C for increased specificity with gene-specific primers
Applications—array labeling, cDNA libraries, RT-PCR, primer extension, 3´ and 5´ RACE

Source
Purified from E. coli expressing the pol gene of M-MLV, mutagenized to increase thermal stability and half-life and to reduce 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® III 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 a 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: 50° C
Final Product Size(s): 12.3 kb or less
Reverse Transcriptase: SuperScript® III
Ribonuclease H Activity: Reduced
Concentration: 200 U⁄µl
Enzyme Function: RNA dependent DNA Polymerase
Form: Frozen
Format: Tube(s)
Number of Reactions: 10 Reactions
Product Size: 2,000 units
Quantity: 2000 units
Sensitivity: Medium
Shipping Condition: Dry Ice
Template: ssRNA

Contents & storage

SuperScript® III Reverse Transcriptase (2,000 units total, at 200U/µL) is supplied with a vial (1 mL) of 5X first-strand buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2], and a vial (500 µL) of 100 mM DTT. Store at -20°C.

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