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VíceDescription:
QuantumScript™ HD Reverse Transcriptase is an engineered version with increased sensitivity, improved specificity and maximum thermostability. QuantumScript™ HD Reverse Transcriptase has been engineered to have longer half life at 50°C, which enables its ability to process longer RNA with more complexed secondary structures. Enhanced thermostability of this enzyme is obtained through re-engineered RNA-based DNA Polymerase domain and the fusion of a novel RNA-interacting surface domain at the RNase H domain site. The enzyme is purified to homogeneity to ensure high thermostability, specificity, fidelity, yield, and more full length cDNA synthesis that the premium reverse transcriptase provides. The optimal fist-strand cDNA synthesis temperature for this enzyme is 50°C, and it has a broad working temperature range from 37° C to 55° C, with cDNA product size from 100 bp to 12 Kb.
Catalog No.
SSIII-100, SSIII-200 and SSIII-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 1 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 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.