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 |
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| 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 |
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| 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
