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SuperScript® III Reverse Transcriptase

日期: 2015-05-26
浏览次数: 105

货号: 18080044

 

规格

 

Optimal Reaction Temperature: 50° C
Final Product Size(s): 12.3 kb or less
Reverse Transcriptase: SuperScript® III
Ribonuclease H Activity: Reduced
Form: Frozen
Format: Tube(s)
Quantity: 10,000 units
Template: ssRNA
Sensitivity: Medium
Product Size: 10,000 units
Concentration: 200 U⁄µl
Enzyme Function: RNA dependent DNA Polymerase
Shipping Condition: Dry Ice
Number of Reactions: 50 Reactions

 

内容及储存

 

SuperScript® III Reverse Transcriptase 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.

 

描述

 

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 generating cDNA from total or poly (A)+ RNA sample. Like SuperScript® II, it synthesizes a complementary DNA strand from single-stranded RNA, DNA, or 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, and 3´ and 5´ RACE

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

Performance and Quality Testing: SDS-PAGE purity; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays (Figure 2) 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 (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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