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Reverse Transcription (cDNA Synthesis)
Product Listing
Application Overview
The synthesis of DNA from an RNA template, via reverse transcription, produces complementary DNA (cDNA). Reverse transcriptases (RTs)
use an RNA template and a short primer complementary to the 3' end of the RNA to direct the synthesis of the first strand cDNA, which can
be used directly as a template for the Polymerase Chain Reaction (PCR). This combination of reverse transcription and PCR (RT-PCR)
allows the detection of low abundance RNAs in a sample, and production of the corresponding cDNA, thereby facilitating the cloning of low
copy genes. Alternatively, the first-strand cDNA can be made double-stranded using DNA Polymerase I and DNA Ligase. These reaction
products can be used for direct cloning without amplification. In this case, RNase H activity, from either the RT or supplied exogenously, is
required.
Many RTs are available from commercial suppliers. Avian Myeloblastosis Virus (AMV) Reverse Transcriptase and Moloney Murine Leukemia
Virus (M-MuLV, MMLV) Reverse Transcriptase are RTs that are commonly used in molecular biology workflows. M-MuLV Reverse
Transcriptase lacks 3´ → 5´ exonuclease activity. 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 50°C, providing higher specificity, higher yield of cDNA and more full-length cDNA product, up
to 12 kb in length.
The use of engineered RTs improves the efficiency of full-length product formation, ensuring the copying of the 5' end of the mRNA transcript
is complete, and enabling the propagation and characterization of a faithful DNA copy of an RNA sequence. The use of the more
thermostable RTs, where reactions are performed at higher temperatures, can be very helpful when dealing with RNA that contains high
amounts of secondary structure.
For help comparing the RTs and cDNA Synthesis reagents available, view our RT/cDNA Synthesis selection chart.
Featured Products
ProtoScript® II Reverse Transcriptase
ProtoScript® First Strand cDNA Synthesis Kit
See entire list »
Protocols
Legal Information
Protocols for Reverse Transcription (cDNA
Synthesis)
First Strand cDNA Synthesis (Standard Protocol) (NEB #M0277)
First Strand cDNA Synthesis (No-RT Negative Control Reaction) (NEB #M0277)
First Strand cDNA Synthesis (No-RT Negative Control Reaction) (NEB #M0253)
First Strand cDNA Synthesis (No-RT Negative Control Reaction) (NEB #M0368)
First Strand cDNA Synthesis (Quick Protocol) (NEB #M0253)
First Strand cDNA Synthesis (Quick Protocol) (NEB #M0277)
First Strand cDNA Synthesis (Quick Protocol) (NEB #M0368)
First Strand cDNA Synthesis (Standard Protocol) (NEB #M0253)
First Strand cDNA Synthesis (Standard Protocol) (NEB #M0368)
First Strand cDNA Synthesis Protocols (E6300)
First Strand cDNA Synthesis Protocols (E6560)
Poly(A) Tailing of RNA using E. coli Poly(A) Polymerase (NEB# M0276)
Legal and Disclaimers
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New
England Biolabs, Inc (NEB).
While NEB develops and validates its products for various applications, the use of this product may require
the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at
[email protected].
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.
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