Neb reverse transcriptase kit

ProtoScript® II Reverse Transcriptase is a recombinant M-MuLV reverse transcriptase with reduced RNase H activity and increased thermostability. It can be used to synthesize first strand cDNA at higher temperatures than the wild type M-MuLV. The enzyme is active up to 48°C, providing higher specificity, higher yield of cDNA and more full-length cDNA product up to 12 kb.

Protoscript II Reverse Transcriptase performs as well as other RNase H– Reverse Transcriptases Robust cDNA synthesis is achieved even with longer templates Generate high quality cDNA even with very low amounts of starting RNA ProtoScript II Reverse Transcriptase displays superior sensitivity

Product Source

The gene encoding a mutant M-MuLV Reverse Transcriptase (RNase H – ) is expressed in E. coli and purified to near homogeneity.

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
ProtoScript ® II Reverse Transcriptase M0368SVIAL -20 1 x 0.02 ml 200,000 units/ml
ProtoScript ® II Reverse Transcriptase Reaction Buffer B0368SVIAL -20 1 x 1.5 ml 5 X
DTT B1034AVIAL -20 1 x 0.5 ml 0.1 M
ProtoScript ® II Reverse Transcriptase M0368LVIAL -20 1 x 0.05 ml 200,000 units/ml
ProtoScript ® II Reverse Transcriptase Reaction Buffer B0368SVIAL -20 1 x 1.5 ml 5 X
DTT B1034AVIAL -20 1 x 0.5 ml 0.1 M
ProtoScript ® II Reverse Transcriptase M0368LVIAL -20 4 x 0.05 ml 200,000 units/ml
DTT B1034AVIAL -20 4 x 0.5 ml 0.1 M
ProtoScript ® II Reverse Transcriptase Reaction Buffer B0368SVIAL -20 4 x 1.5 ml 5 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 1 nmol of dTTP into acid-insoluble material in a total reaction volume of 50 μl in 10 minutes at 37°C using poly(rA)•oligo(dT)18 as template.

Reaction Conditions

1X ProtoScript ® II Reverse Transcriptase Reaction Buffer
Incubate at 42°C

1X ProtoScript ® II Reverse Transcriptase Reaction Buffer
50 mM Tris-HCl
75 mM KCl
3 mM MgCl2
(pH 8.3 @ 25°C)

Storage Buffer

20 mM Tris-HCl
100 mM NaCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.01% IGEPAL® CA-630
pH 7.5 @ 25°C

Heat Inactivation

65°C for 20 minutes

Unit Assay Conditions

50 mM Tris-HCl (pH 8.3), 75 mM KCl, 6 mM MgCl2, 10 mM dithiothreitol, 0.01% IGEPAL CA-630, 0.5 mM dTTP, 0.4 mM poly(rA)•oligo(dT)18.

Companion Products
Notes
References
Protocols
  1. First Strand cDNA Synthesis (Quick Protocol) (NEB #M0368)
  2. First Strand cDNA Synthesis (Standard Protocol) (NEB #M0368)
  3. First Strand cDNA Synthesis (No-RT Negative Control Reaction) (NEB #M0368)
Usage Guidelines
Selection Charts
FAQs
Citations

Additional Citations
Quality Control Assay

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications
Certificate of Analysis
Safety Data Sheets
Legal And Disclaimer

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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.