Reverse Transcriptase Selection Chart

Reverse Transcriptase Maximum RT Product Length  Recommended Reaction Temperature Active Temperature Range Heat Inactivation RNase H Activity Strand Displacement Can the RT add adaptors to 5′ RNA/3′ cDNA end? WarmStart® Aptamer Formats Available **** Applications & Features
ProtoScript® II RT 12kb 42°C 37°C-48°C 80°C/ 5min - * + -  ** No ProtoScript® II Reverse Transcriptase Ideal for first strand synthesis of targets up to 12kb, endpoint RT-PCR
ProtoScript® II First Strand cDNA Synthesis Kit
OneTaq® One-Step RT-PCR Kit
Luna® RT 3kb *** 55°C 55°C-65°C 95°C/ 1min - * + -  ** No LunaScript® RT SuperMix

Ideal for two step RT-qPCR, first-strand synthesis, endpoint RT-PCR, and amplicon sequencing

Single-tube 5X supermix contains everything needed for cDNA synthesis except your template

LunaScript® RT Master Mix Kit (Primer-free) Single-tube 5X master mix containing all the necessary components for first strand cDNA synthesis except primers. Includes a separate No-RT Control Mix.
LunaScript® RT SuperMix Kit LunaScript RT SuperMix with No-RT Control and nuclease-free water components
Yes Luna® Universal One-Step RT-qPCR Kit Ideal for dye-based one-step RT-qPCR
Luna® Universal Probe One-Step RT-qPCR Kit Ideal for probe-based one-step RT-qPCR
Luna® Probe One-Step RT-qPCR Kit (No ROX) Mix does not include universal ROX reference dye
Luna® Probe One-Step RT-qPCR 4X Mix with UDG

4X mix for increased sample input

Inclusion of UDG (and dUTP) prevents carryover contamination

LyoPrime Luna® Probe One-Step RT-qPCR Mix with UDG Lyophilized format
LunaScript® Multiplex One-Step RT-PCR Kit

Endpoint PCR from RNA targets using Q5® High-Fidelity DNA Polymerase

WarmStart® RTx 5kb 65°C
(RT-LAMP)
45°C-65°C 80°C/ 10min + + -  ** Yes WarmStart® RTx Reverse Transcriptase Ideal for RT-LAMP, cDNA synthesis
55°C
(cDNA synthesis)
WarmStart® Colorimetric LAMP 2X Master Mix (DNA & RNA) Colorimetric LAMP mixes permit a simple readout with visible color change
WarmStart® Colorimetric LAMP 2X Master Mix with UDG The inclusion of UDG (and dUTP) prevents carryover contamination
WarmStart® LAMP Kit (DNA & RNA) Non-colorimetric LAMP mixes enable multiple readouts and buffered samples
WarmStart® Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) The inclusion of UDG (and dUTP) prevents carryover contamination
WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) Kit includes Multi-Purpose LAMP Master Mix and 50X fluorescent dye
Reverse Transcriptase Maximum RT Product Length  Recommended Reaction Temperature Active Temperature Range Heat Inactivation RNase H Activity Strand Displacement Can the RT add adaptors to 5′ RNA/3′ cDNA end? WarmStart® Aptamer Formats Available **** Applications & Features
Template Switching RT 10-12kb 42°C 37°C-48°C 85°C/ 5min - * + +++ 1 No Template Switching RT Enzyme Mix Ideal for 5′ RACE, template-switching cDNA amplification, second-strand synthesis
Induro® RT >20kb 55°C 45°C-60°C 95°C/ 1min - +++ 2 -  ** No Induro® Reverse Transcriptase

Fast, processive, intron-encoded RT

Ideal for long transcripts, direct RNA sequencing, cDNA synthesis from samples with strong secondary structures and/or with inhibitors

AMV RT 5kb 42°C 37°C-52°C 85°C/ 5min + + not tested No AMV Reverse Transcriptase First-strand synthesis
M-MuLV RT 5kb 42°C 37°C-45°C 65°C/ 20min + + + ** No M-MuLV Reverse Transcriptase First-strand synthesis
ProtoScript® First Strand cDNA Synthesis Kit Kit includes RT, oligo-dT primers, randomized primer mix, and nuclease-free water

* Engineered to have an inactive RNase H domain, but does still possess some RNase H activity.
** The RT does possess terminal transferase activity, but the added nontemplated nucleotides are not suitable for efficient adaptor ligation by template switching.
*** Using random hexamers and poly-d(T) primers. Using gene-specific primers can yield up to 12kb cDNA products. Our one-step RT-qPCR Luna® mixes are formulated to produce cDNA less than 1kb in length.
**** Bulk, custom, and glycerol-free formats are available. Please visit www.neb.com/glycerol-free for more info.


1 Wulf et al. (2019) J Biol Chem. 294(48), 18220-18231). PMID: 31640989
2 Maguire and Guan (2022) PLOS ONE 17(10) e0275471. PMID: 36215256