DNA Amplification Reagents - Reverse transcriptase

SuperScript® II reverse transcriptase DNase, RQ1, RNase-free source: bovine pancreas

CP RQ1 RNase-free DNase is used in DNase treatment of 32 RNA samples prior to RT-PCR, preparation of DNA-free RNA, degradation of DNA from RNA transcription SuperScript® II reverse transcriptase (RT) is an improved version of SuperScript® RT. systems, nick translation of DNA, and studying DNA- protein interactions by DNase I footprinting. DNA polymerase that synthesises a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. This is genetically engineered by the RQ1 RNase-free DNase is a preparation of I that degrades single- introduction of point mutations rather than a deletion in the RNase H active center. Like stranded or double-stranded DNA to produce 3’-hydroxyl oligonucleotides. This preparation SuperScript® RT, SuperScript® II RT has reduced RNase H activity. Unlike SuperScript® RT, is qualified for use in applications where maintaining the integrity of RNA is critical. The however, the selective mutations within the RNase H domain maintain full polymerase absence of detectable RNase activity is determined by two different methods including gel activity. This structural modification eliminates degradation of RNA molecules during first- analysis of a mixture of RNA and DNA. strand cDNA synthesis and gives SuperScript® II RT superior performance characteristics, including: Catalogue No Quantity Price BPE3223-1 1,000 units 42.60 • Greater first-strand cDNA yields • More full length cDNA synthesis Provided with 10X reaction buffer: 400mM Tris-HCl (pH8.0), 100mM MgSO4, and 10mM • Full activity at 42°C CaCl2. One unit is defined as the amount of enzyme required to completely degrade 1µg of DNA Source: Purified from E. coli expressing the pol gene of M-MLV (1,2), mutagenised to in 10min at 37°C in 50µL of a buffer containing 40mM Tris-HCl (pH7.9), 10mM NaCl, 6mM reduce the RNase H activity. MgCl2, and 10mM CaCl2

Performance and quality testing: SDS-PAGE purity; , Concentration: 1unit/µL , and assays; and yield and length of cDNA product. Storage buffer: 10mM HEPES (pH7.5), 50% (v/v) glycerol, 10mM CaCl , and 10mM MgCl 2 2 Unit definition: One unit of SuperScript® II is the amount of enzyme required to Product specification incorporate 1nmole of deoxyribonucleotide into acid-precipitable material in 10min at 37°C Tested for: Activity, RNase and specific performance test. using poly(A) oligo(dT)12-18 as template primer.

Unit reaction conditions: 50mM Tris-HC1 (pH8.3), 40mM KC1, 6mM MgC12, 1mM DTT, 0.5mM [3H]dTTP, 0.1mM poly(A), 0.1mM oligo(dT)12-18, 0.1mg/m1 BSA, and enzyme in 50µL for 10min. at 37°C.

Catalogue No Description Units Price VX18064022 SuperScript® II reverse transcriptase 2,000 38.50 VX18064014 SuperScript® II reverse transcriptase 10,000 164.00 VX18080085 SuperScript® II reverse transcriptase 4 x 10,000 586.00

SuperScript® III reverse transcriptase

CP

Synthesis of first-strand cDNA, array labelling, cDNA libraries, RT-PCR*, primer extension, and 3’ and 5’ RACE.

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 220min, providing increased specificity with Gene- Specific Primers (GSPs) and the highest cDNA yield of all MoreRTs.

Like SuperScript® II, it synthesises 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. The structural modifications provide:

• Half life of 220min at 50°C for the highest cDNA yields • Reduced RNase H activity for more full-length cDNA • Full activity at 50°C for increased specificity with GSP • Ability to increase RT units without inhibiting subsequent PCR

Source: Purified from E. coli expressing the pol gene of M-MLV (1,2), mutagenised to increase thermal stability and half life and 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 1nmole of deoxyribonucleotide into acid-precipitable material in 10min at 37°C using poly(A) oligo(dT)12-18 as a template primer.

Unit reaction conditions: 50mM Tris-HCl (pH8.3), 40mM KCl, 6mM MgCl2 , 1mM DTT. 0.5 mM [3 H]dTTP, 0.1mM poly(A), 0.1mM oligo(dT)12-18 , 0.1mg/mL BSA, and enzyme in 50µL for 10min at 37°C.

Catalogue No Description Units Price VX18080093 SuperScript® III reverse transcriptase 2,000 40.00 VX18080044 SuperScript® III reverse transcriptase 10,000 166.00 *Polymerase Chain Reaction (PCR) is a process covered by patents owned by Hoffman-La Roche VX18080085 SuperScript® II reverse transcriptase 4 x 10,000 586.00

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