Description:
QuantumScript™ Reverse Transcriptase is a new engineered version of M-MLV reverse transcriptase with a minimum RNase H activity and enhaced thermostability. The enzyme is purified to homogeneity to ensure high performance. The optimal fist-strand cDNA synthesis temperature for this enzyme is 42°C, with cDNA product size from 100 bp to 7 Kb.
Catalog No.
SSII-100, SSII-200 and SSII-300
Source
E.coli
Concentration
200 u/μl
Storage Buffer
20 mM Tris-HCl (pH 7.5), 1 mM DTT, 0.05% (v/v)Triton X-100, 0.1 mM EDTA, 0.1 M NaCl and 50% (v/v) glycerol.
Reaction Buffer (5x)
250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2, and 50 mM DTT
Unit Definition
One unit of the enzyme incorporates I nmole of dTTP into acid-precipitable material in 10 minutes at 37˚C using poly (A): oligo (dT)25 as template-primer.
Quality Control
This enzyme has passed the quality control assays: SDS-PAGE analysis for purity, functional absence of endonuclease/nickase activities, functional absence of exonuclease activities, functional absence of protease activity.
Storage and Handling -20˚C
Protocol
First-Strand cDNA Synthesis
Materials to Be Supplied by the User
The following procedure uses 10 pg to 5 µg of total RNA or 10 pg to 500 ng of mRNA.
Note: The 5X Reaction Buffer is compatible with enzymes used in a number of downstream applications. Typically there is no need for phenol extractions or ethanol precipitations using this protocol before any PCR amplification