Invitrogen™ SuperScript™ III Reverse Transcriptase 

Description

SuperScript III Reverse Transcriptase (RT) 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.

• Thermostability: half-life of 220 minutes 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
• Concentration: 200U/μL

• Unit definition: One unit of SuperScript III RT is the amount of enzyme required to incorporate 1nmole of deoxyribonucleotide into acid-precipitable material in 10 minutes at 37°C using poly(A)·oligo(dT)_12-18 as a template·primer
• Unit reaction conditions: 50mM Tris-HCl (pH 8.3), 40mM KCl, 6mM MgCl₂, 1mM DTT, 0.5mM [³H]dTTP, 0.1mM poly(A), 0.1mM oligo(dT)_12-18, 0.1mg/mL BSA, and enzyme in 50μL for 10 minutes at 37°C
• Optimal reaction temperature: 50°C
• Final product size: 12.3kb or less

PCR and Real-Time PCR, Reverse Transcription, array labeling, cDNA libraries, primer extension, 3' and '5 RACE

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