Reverse Transcriptase Selection Chart
| Product | Maximum RT Product Length | Recommended Reaction Temperature | Active Temperature Range | Heat Inactivation | RNase H Activity | Strand Displacement | WarmStart® Aptamer | Applications & Features | ||
|---|---|---|---|---|---|---|---|---|---|---|
| ProtoScript® II RT | ||||||||||
| ProtoScript® II Reverse Transcriptase | 12 kb | 42°C | 37°C−48°C | 65°C/ 20 min 80°C/ 5 min 95°C/ 1 min |
-* | + | No | Ideal for first strand synthesis of targets up to 12 kb, endpoint RT-PCR | ||
| ProtoScript® II First Strand cDNA Synthesis Kit | 12 kb | 42°C | 37°C−48°C | 65°C/ 20 min 80°C/ 5 min 95°C/ 1 min |
-* | + | No | |||
| OneTaq® One-Step RT-PCR Kit | 12 kb | 42°C | 37°C−48°C | 65°C/ 20 min 80°C/ 5 min 95°C/ 1 min |
-* | + | No | |||
| Luna® RT | ||||||||||
| LunaScript® RT SuperMix | 3 kb ** | 55°C | 55°C−65°C | 95°C/ 1 min | -* | + | No | 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 |
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| LunaScript® RT SuperMix Kit | 3 kb ** | 55°C | 55°C−65°C | 95°C/ 1 min | -* | + | No | LunaScript RT SuperMix without No-RT Control and nuclease-free water components | ||
| LunaScript® RT Master Mix Kit (Primer-free) | 3 kb | 55°C | 55°C−65°C | 95°C/ 1 min | -* | + | No | Single-tube 5X master mix containing all the necessary components for first strand cDNA synthesis except primers Includes a separate No-RT Control Mix |
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| Luna® Universal One-Step RT-qPCR Kit | 3 kb | 55°C | 55°C−65°C | 95°C/ 1 min | -* | + | Yes | Ideal for dye-based one-step RT-qPCR | ||
| Luna® Universal Probe One-Step RT-qPCR Kit | 3 kb | 55°C | 55°C−65°C | 95°C/ 1 min | -* | + | Yes | Ideal for probe-based one-step RT-qPCR | ||
| Luna® Probe One-Step RT-qPCR Kit (No ROX) | 3 kb | 55°C | 55°C−65°C | 95°C/ 1 min | -* | + | Yes | Ideal for probe-based one-step RT-qPCR | ||
| Luna® Probe One-Step RT-qPCR 4X Mix with UDG | 3 kb | 55°C | 55°C−65°C | 95°C/ 1 min | -* | + | Yes | 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 | 3 kb | 55°C | 55°C−65°C | 95°C/ 1 min | -* | + | Yes | Lyophilized format for probe-based one-step RT-qPCR | ||
| LunaScript® Multiplex One-Step RT-PCR Kit | 3 kb | 55°C | 55°C−65°C | 95°C/ 1 min | -* | + | Yes | Endpoint PCR from RNA targets using Q5® High-Fidelity DNA Polymerase | ||
| WarmStart® RTx | ||||||||||
| WarmStart® RTx Reverse Transcriptase | 5 kb | 65°C (RT-LAMP) 55°C (cDNA synthesis) |
45°C−65°C | 80°C/ 10 min 95°C/ 1 min |
+ | + | Yes | Ideal for RT-LAMP | ||
| WarmStart® Colorimetric LAMP 2X Master Mix (DNA & RNA) | 5 kb | 65°C (RT-LAMP) 55°C (cDNA synthesis) |
45°C−65°C | 80°C/ 10 min 95°C/ 1 min |
+ | + | Yes | Colorimetric LAMP mixes permit a simple readout with visible color change | ||
| WarmStart® Colorimetric LAMP 2X Master Mix with UDG | 5 kb | 65°C (RT-LAMP) 55°C (cDNA synthesis) |
45°C−65°C | 80°C/ 10 min 95°C/ 1 min |
+ | + | Yes | Colorimetric LAMP mix. The inclusion of UDG (and dUTP) prevents carryover contamination | ||
| WarmStart® LAMP Kit (DNA & RNA) | 5 kb | 65°C (RT-LAMP) 55°C (cDNA synthesis) |
45°C−65°C | 80°C/ 10 min 95°C/ 1 min |
+ | + | Yes | Non-colorimetric LAMP mix enables multiple readouts and buffered samples | ||
| WarmStart® Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) | 5 kb | 65°C (RT-LAMP) 55°C (cDNA synthesis) |
45°C−65°C | 80°C/ 10 min 95°C/ 1 min |
+ | + | Yes | LAMP Mix that is compatible with multiple detection methods. The inclusion of UDG (and dUTP) prevents carryover contamination | ||
| WarmStart® Fluorescent LAMP/RT-LAMP Kit (with UDG) | 5 kb | 65°C (RT-LAMP) 55°C (cDNA synthesis) |
45°C−65°C | 80°C/ 10 min 95°C/ 1 min |
+ | + | Yes | Kit includes Multi-Purpose LAMP Master Mix and 50X fluorescent dye | ||
| Template Switching RT | ||||||||||
| Template Switching RT Enzyme Mix | 10-12 kb | 42°C | 37°C−48°C | 80°C/ 5 min | - * | + | No | Ideal for 5′ RACE, template-switching cDNA amplification, second-strand synthesis Can add adaptors to 5′ RNA/3′ cDNA end |
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| Induro® RT | ||||||||||
| Induro® Reverse Transcriptase | >20 kb | 55°C | 45°C−60°C | 95°C/ 1 min | - | +++ 2 | No | Fast, processive, intron-encoded RT Ideal for long transcripts, direct RNA sequencing, cDNA synthesis from samples with strong secondary structures and/or with inhibitors |
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| AMV RT | ||||||||||
| AMV Reverse Transcriptase | 5 kb | 42°C | 37°C−50°C | 80°C/ 5 min 95°C/ 1 min |
+ | + | No | First-strand synthesis | ||
| M-MuLV RT | ||||||||||
| M-MuLV Reverse Transcriptase | 5 kb | 42°C | 37°C−45°C |
65°C/ 20 min 90°C/ 5 min 95°C/ 1 min |
+ | + | No | First-strand synthesis Can add adaptors to 5′ RNA/3′ cDNA end but not suitable for efficient adaptor ligation by template switching |
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| ProtoScript® First Strand cDNA Synthesis Kit | 10 kb | 42°C | 37°C−45°C | 80°C/ 5 min | + | + | No | 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.
** 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
