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. 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, may be helpful for reverse transcription of RNA that containing secondary structure.
For help comparing the RTs and cDNA Synthesis reagents available, view our RT/cDNA Synthesis selection chart.
- First Strand cDNA Synthesis Protocols (E6300)
- First strand cDNA synthesis OneTaq® RT-PCR Kit
- PCR Amplification with OneTaq® RT-PCR Kit
- First Strand cDNA Synthesis (Quick Protocol) (NEB #M0277)
- First Strand cDNA Synthesis Protocols (E6560)
- A-Tailing with Klenow Fragment (3'→5' exo-)
- Protocol for blunting ends by 3' overhang removal and fill-in of 3' recessed (5' overhang) ends using DNA Polymerase I, Large (Klenow) Fragment (M0210)
- Protocol for Avoiding Rnase Contamination using Rnase Inhibitor, Human Placenta (M0307)
- Loop-mediated Isothermal Amplification (LAMP)
- One-Step RT-PCR Protocols (E5315)
- Reaction Conditions (E5315)
- Standard PCR Protocol (E5315)
- Typical cDNA Synthesis Protocol with WarmStart RTx (NEB #M0380)
- Typical RT-LAMP Protocol
- First Strand cDNA Synthesis (No-RT Negative Control Reaction) (NEB #M0277)
- First Strand cDNA Synthesis (Standard Protocol) (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 #M0368)
- First Strand cDNA Synthesis (Standard Protocol) (NEB #M0253)
- First Strand cDNA Synthesis (Standard Protocol) (NEB #M0368)
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