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Reverse Transcriptase (cDNA Synthesis)

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Introduction to Reverse Transcriptase (RT) Enzymes

The enzyme reverse transcriptase originates from viruses, mainly retroviruses. The RT enzyme generates complementary DNA (cDNA) from RNA templates. On this website a number of RT enzymes are listed, most are based on the M-MLV (moloney murine leukemia virus) or on AMV (avian myeloblastosis virus) RT enzymes.

Some provider modified the enzymes in order to alter enzyme characteristics. Retroviral RT enzymes possess different enzymatic functions and properties. Some of the introduced kits lack the RNase H activity. The RNase H activity degrades the template during long incubation time, kits that lack this activity are especially useful for generation of long cDNAs. Long cDNAs are important for long read sequencing as performed with third generation sequencing technologies from PacBio and Oxford Nanopore.

The usage of highly purified or cloned and modified enzymes result in improve enzyme activity and sensitivity. The modification to more thermostable enzymes enables higher specific reactivity. These Enzymes allow performing of reverse transcription at 42°C or even at higher temperatures.

This website also introduces kits that include specific buffer and protocols in order to enable cDNA synthesis without prior RNA purification. You will find also kits that enable cDNA syntheses from total RNA, from polyA RNA (mRNA) or from micro RNA. The generation of cDNA from total RNA is for example important for transcriptome sequencing and sequencing of microbiome that contain viruses.


All-in-One™ First-Strand cDNA Synthesis Kit

The All-in-One™ First-Strand cDNA Synthesis Kit includes a reverse transcriptase and a specialized set of reagents designed to yield cDNA that is optimal for gene cloning, cDNA library creation and quantitative PCR amplification. The kit uses Moloney Murine Leukemia Virus Reverse Transcriptase, RNase H Minus (M-MLV RT (H–)) which is an RNA-dependent DNA polymerase that is used in cDNA synthesis with long RNA templates. The lack of RNase H activity is important in this application in that RNase H activity will start to degrade template during long incubation times which are required for producing long cDNAs. RNase H minus RT enables preparation of long cDNAs and libraries containing a high percentage of full-length cDNA.
Source: Gene Copoeia

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AMPIGENE® cDNA Synthesis Kit Enzo

AMPIGENE® cDNA Synthesis Kit uses the latest developments in reverse transcriptase (RTase) technology and buffer chemistry for efficient and accurate cDNA synthesis.
The modified MMLV RTase is blended with RNase inhibitor to prevent degradation of RNA by contaminating RNase. The RTase is thermostable and extremely active.

Source: Enzo Lifesciences

AMV RT

Avian Myeloblastosis Virus Reverse Transcriptase (AMV RT) catalyzes the polymerization of DNA using template DNA, RNA or RNA:DNA hybrids.

Source: Promega

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Cells-to-cDNA™ II Kit Ambion

The Ambion® Cells-to-cDNA™ II Kit (patent pending) produces cDNA from cultured mammalian cells in less than 2 hr. No RNA isolation is required. This kit contains sufficient reagents for 40 reactions and is ideal for those who want to perform reverse transcription reactions on small numbers of cells, numerous cell samples, or for scientists who are unfamiliar with RNA isolation.

Source: Thermo Fisher

CLONED AMV FIRST-STRAND CDNA SYNTHESIS KIT INVITROGEN

The Cloned AMV First-Strand Synthesis Kit combines the high performance of Invitrogen™ Cloned AMV RT with the convenience of optimized, pre-qualified reagents. The and ! subunits of the avian myeloblastosis virus gene were co-expressed and purified to produce this recombinant AMV RT. Unlike the commonly available native enzyme, Invitrogen™ Cloned AMV is a high-purity preparation that results in greater specific activity, leading to improved yields and superior sensitivity. To achieve high-quality routine RT-PCR, combine with Platinum® Taq DNA Polymerase for high-specificity PCR or with Platinum® Taq DNA Polymerase High Fidelity for subsequent PCR cloning.

Source: Thermo Fisher

Enhanced Avian First Strand Synthesis Kit Sigma Aldrich

Enhanced Avian First Strand Synthesis Kit utilizes a highly purified avian myeloblastosis virus reverse transcriptase (eAMV-RT) that offers superior performance in comparison to standard AMV-RT or standard Moloney murine leukemia virus reverse transcriptase (MMLV-RT). This exceptionally robust eAMV-RT has an enhanced ability to transcribe through difficult secondary structure at elevated temperatures (up to 65 °C) making it the ideal enzyme for producing high quality full-length cDNA from total RNA or poly(A)+ RNA.

Source: Sigma Aldrich

Enhanced Avian Reverse Transcriptase [eAMV™ RT]

Enhanced Avian Reverse Transcriptase (eAMV RT) is used to transcribe RNA into DNA, and facilitates efficient mRNA template driven sysnthesis of cDNAs. This is due to the abillity of this enhanced AMV-RT to transcribe large mRNA templates, to transcribe through difficult secondary structures, and to detect low abundance mRNAs by RT-PCR.

Source: Sigma Aldrich

EnzScript™ (MMLV Reverse Transcriptase RNase H-)

EnzScript™ (MMLV Reverse Transcriptase RNase H-)
RNA-dependent DNA polymerase with no detectable RNase H activity.

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EpiNext™ Hi-Fi cDNA Synthesis Kit

The EpiNext™ Hi-Fi cDNA Synthesis Kit is a complete set of optimized reagents to synthesize a DNA copy specifically using bisulfite-converted RNA and enriched RNA fragments.

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FastLane Cell cDNA Kit Qiagen

The FastLane Cell cDNA Kit provides a fast and simple procedure for preparing first-strand cDNA directly from cultured cells without RNA purification. The kit is supplied with wash and lysis buffers for preparing lysates and stabilizing RNA, gDNA Wipeout Buffer for eliminating genomic DNA contamination, and all reaction components for fast and efficient cDNA synthesis. The FastLane Cell cDNA Kit is ideal for experiments requiring rapid, high-throughput gene expression analysis.

Source: Qiagen

First Strand cDNA Synthesis Kit for RT-PCR (AMV) Roche

The First Strand cDNA Synthesis Kit is used for the synthesis of the first strand cDNA as the starting reaction for two-step RT PCR. It can be used with either sequence-specific primers, poly(dT)15 primers, or random primers.
The kit includes Reverse Transcriptase AMV for first strand synthesis, two different primers, our PCR Nucleotide Mix, and Control Neo pa RNA. PCR products that are generated by RT-PCR can be cloned using standard procedures.
The use of AMV RT for first-strand cDNA synthesis has certain advantages. The higher thermostability of this enzyme compared to M-MuLV RT allows the reaction to be performed at +42°C, giving higher specificity and better resolution of secondary structures compared to a reaction performed at +37°C.

Source: Roche

GoScript™ Reverse Transcription System

Efficiently transcribe RNA into full-length cDNA. This system is highly resistant to possible contaminants, allowing successful confirmatory testing.

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High-Capacity RNA-to-cDNA™ Kit Applied Biosystems

The High Capacity RNA-to-cDNA™ Kit is a streamlined reverse transcription kit designed for optimum performance with TaqMan® Gene Expression Master Mix, Power SYBR® Green PCR Master Mix, and other PCR enzymes.

Source: Thermo Fisher

ImProm-II™ Reverse Transcription System Promega

The ImProm-II™ Reverse Transcription System produces efficient, robust synthesis of first-strand cDNA in preparation for PCR amplification. The components of the ImProm-II™ Reverse Transcription System can be used to reverse transcribe RNA templates starting with total RNA, poly(A)+ mRNA or synthetic transcript RNA. The optimized reaction buffer and powerful ImProm-II™ Reverse Transcriptase provided in the ImProm-II™ System together enable robust, full-length cDNA synthesis for the reproducible analysis of rare or long messages.

Source: Promega

iScript™ cDNA Synthesis Kit

The iScript cDNA synthesis kit is a sensitive and easy-to-use first-strand cDNA synthesis kit for gene expression analysis using real-time qPCR. This two-tube kit is optimized to yield sensitive, unbiased representation over a broad dynamic range, with minimal setup and reaction time.

Source: Bio-Rad

Maxima™ reverse transcriptase

Thermo Scientific Maxima Reverse Transcriptase (RT) was developed through in vitroevolution of M-MuLV RT. The enzyme possesses an RNA-dependent and DNA-dependent polymerase activity as well as RNase H activity. The engineered enzyme features dramatically improved thermostability and robustness, and increased synthesis rates compared to wild type M-MuLV RT.
Source: ThermoFisher

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MessageSensor™ RT Kit

For the reverse transcription of RNA. The Ambion® kit includes a M-MLV RT that maintains full RNase H activity for increased sensitivity of detection in RT-PCR experiments. Includes sufficient reagents for 50 reactions.
MessageSensor™ is designed to provide unsurpassed results in the least amount of time. The kit contains a unique formulation of M-MLV RT developed for a single tube approach increasing lower-end reaction sensitivity (see figure). The reverse transcription step takes only 15 minutes.

Source: Thermo Fisher

Methylamp H Minus M-MLV Reverse Transcriptase

Methylamp H Minus M-MLV Reverse Transcriptase is a genetically modified M-MLV RT which exhibits RNA or DNA dependent DNA polymerase, but lacks ribonuclease H activity. This enzyme can synthesize a complementary DNA strand initiating from a primer using RNA or DNA templates. Removal of the RNase H activity results in an increase of full-length cDNA products. The enzyme has RNA polymerization-dependent and DNA polymerization-dependent activity but lacks ribonuclease H activity.
Source: Epigentec

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M-MLV Reverse Transcriptase, RNase H Minus, Point Mutant Product Promega

Moloney Murine Leukemia Virus Reverse Transcriptase, RNase H Minus, Point Mutant (M-MLV RT (H–)) is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long messenger RNA templates (>5kb). The lack of RNase H activity is germane to the applications in that this activity can start to degrade templates and first-strand products when incubation times are long, as they may need to be when making long cDNAs. Although many researchers are successful in using M-MLV RT (H+) for analytical and some preparative cDNA applications, reverse transcriptases lacking RNase H activity provide another option for the preparation of long cDNAs and libraries containing a high percentage of full-length cDNA.

Source: Promega

MultiScribe™ Reverse Transcriptase

MultiScribe™ Reverse Transcriptase is a recombinant moloney murine leukemia virus (rMoMuLV) reverse transcriptase that has been optimized for TaqMan-based assays. MultiScribe™ Reverse Transcriptase is similar to MuLV reverse transcriptase but differs in its recommended usage. MultiScribe™ Reverse Transcriptase is a recombinant RNA–dependent DNA polymerase that uses single-stranded RNA as a template in the presence of a primer to synthesize a complementary DNA (cDNA) strand.
Source: Thermo Fisher

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MystiCq® microRNA cDNA Synthesis Mix Sigma Aldrich

Starting with total RNA or samples pre-enriched or microRNAs, the MystiCq microRNA cDNA Synthesis Mix kit contains all the components necessary to convert mature microRNAs into cDNA templates for qPCR. Since microRNAs are not naturally polyadenylated, the first step in the cDNA synthesis process is to polyadenylate the microRNAs through a poly(A) polymerase reaction. Next, the ReadyScript reverse transcriptase, an oligo-dT adapter primer and other required reagents are added to convert the polyadenylated microRNAs into cDNA. The adapter primer includes a unique sequence at the 5’ end that is complementary to the sequence of the MystiCq Universal PCR Primer, another component of the MystiCq microRNA qPCR Assay System.The kit also contains a Positive Control Primer that is specific to SNORD44, a small nucleolar RNA that is widely expressed in most human tissues.

Source: Sigma Aldrich

NxGen® M-MuLV Reverse Transcriptase

NxGen® M-MuLV Reverse Transcriptase is an RNA-dependent DNA polymerase which shows no measurable 3′→5′ proofreading function. This enzyme can copy a single-stranded DNA template or perform cDNA synthesis by extending a DNA primer annealed to an RNA template.
Source: lucigen

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Omniscript RT Kit Print Qiagen

The Omniscript Reverse Transcription (RT) Kit is specially designed for reverse transcription with any amount of RNA from 50 ng to 2 μg per reaction. A high affinity for RNA allows Omniscript Reverse Transcriptase to provide superior performance compared with other reverse transcriptases, delivering higher sensitivity in RT-PCR, even with low-copy numbers. The Omniscript RT Kit includes Omniscript Reverse Transcriptase, dNTPs and an optimized reaction buffer. Simply add primers for fast and easy cDNA synthesis.

Source: Qiagen

Ovation RNA-Seq System V2

The Ovation RNA-Seq System V2 provides a rapid, simple and sensitive RNA amplification process for preparing amplified cDNA from total RNA for RNA-Seq applications. Amplification is initiated at the 3' end as well as randomly throughout the whole transcriptome in the sample making the Ovation RNA-Seq System V2 ideal for amplification of both mRNA and non-polyadenylated transcripts. The Ovation RNA-Seq System V2 is powered by Ribo-SPIA® technology, a rapid and sensitive amplification process developed by NuGEN.
Source: NuGEN

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Ovation RNA-Seq FFPE System

The Ovation RNA-Seq FFPE System provides a fast and simple method for preparing amplified cDNA from FFPE-derived total RNA for RNA-Seq applications (transcriptome sequencing). Amplification is initiated at the 3' end as well as randomly throughout the transcriptome in the sample, making the Ovation RNA-Seq FFPE System ideal for amplification of the severely degraded and chemically modified RNA typically obtained from FFPE samples.
Source: NuGEN

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Ovation RNA Archiving - cDNA Module

The Ovation RNA Archiving - cDNA Module is a custom solution for fast and simple generation of multiple aliquots of ds- cDNA from as little as 20 ng total RNA input. The resulting ds-cDNA product is stable at -20C, while the original sample material is preserved. The minimal cost associated with the generation of SPIA tagged ds-cDNA makes it amenable for use with all incoming samples.
Source: NuGEN

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PrimeScript Reverse Transcriptase

PrimeScript Reverse Transcriptase has exceptionally strong strand displacement activity and enables efficient preparation of cDNA up to 12 kb in length. It is robust, versatile and well-suited for applications requiring full-length cDNA such as preparation of cDNA libraries and other techniques involving first strand cDNA synthesis (RT-PCR, preparation of cDNA probes, real-time quantitative RT-PCR).
Source: Clonetech

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ProtoScript® II Reverse Transcriptase

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 48°C, providing higher specificity, higher yield of cDNA and more full-length cDNA product up to 12 kb.
NEB offers several reagents for cDNA Synthesis for use in applications including qPCR and qRT-PCR. Reagents are available as kits or standalone products.
Source: NEB

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QIAGEN OneStep RT-PCR Kit

The QIAGEN One-Step RT-PCR Kit provides a blend of Sensiscript and Omniscript Reverse Transcriptases, HotStarTaq DNA Polymerase, QIAGEN OneStep RT-PCR Buffer, a dNTP mix, and Q-Solution, a novel additive that enables efficient amplification of "difficult" (e.g., GC-rich) templates. The easy one-tube setup and optimized components result in highly sensitive and successful results.

Source: Qiagen

QuantiTect Reverse Transcription Kit Qiagen

The unique QuantiTect Reverse Transcription Kit provides a fast and convenient procedure for cDNA synthesis with integrated genomic DNA removal. Genomic DNA contamination in RNA samples is effectively eliminated by gDNA Wipeout Buffer. All components that are required for fast and efficient reverse transcription are provided with the QuantiTect Reverse Transcription Kit, including Quantiscript Reverse Transcriptase, Quantiscript RT Buffer, and a unique RT Primer Mix. The synthesized cDNA is optimized for use in real-time PCR, allowing reliable quantification of targets from all regions of mRNA transcripts.

Source: Qiagen

QUANTITECT WHOLE TRANSCRIPTOME KIT

The QuantiTect Whole Transcriptome Kit enables the preamplification and reverse transcription of limited amounts of RNA to high yields of cDNA for unlimited gene expression analysis using real-time PCR. An optimized protocol ensures a fast and easy procedure. The kit consists of a complete set of enzymes and buffers for whole transcriptome amplification (WTA). Innovative modification of Phi 29 polymerase technology allows yields of up to 40 μg cDNA from as little as 1 ng RNA. The uniquely high processivity of this polymerase guarantees the generation of cDNA containing uniformly amplified targets to ensure reliable gene expression analysis in real-time PCR.

Source: Qiagen

ReadyScript® cDNA Synthesis Mix Sigma Aldrich

ReadyScript cDNA Synthesis Mix is a sensitive and easy-to-use solution for two-step RT-PCR. This 5X concentrated mix provides all necessary components (except RNA template) for first-strand synthesis including: buffer, dNTPs, MgCl2, primers, RNase inhibitor protein, ReadyScript reverse transcriptase and stabilizers. ReadyScript is a RNase H(+) derivative of MMLV reverse transcriptase, optimized for reliable cDNA synthesis over a wide dynamic range of input RNA. The unique blend of oligo (dT) and random primers in the ReadyScript cDNA Synthesis Mix works exceptionally well with a wide variety of targets. It is optimized for the production of targets < 1kb in length. ReadyScript cDNA Synthesis Mix produces excellent results in both real-time and conventional RT-PCR.

Source: Sigma Aldrich

Ready-To-Go T-Primed First-Strand Kit

The kit provides 50 single-dose, ambient-temperature-stable, first-strand cDNA synthesis reactions, each containing NotI-(dT)18 Primer, Cloned FPLCpure M-MuLV Reverse Transcriptase, RNase Inhibitor, and nucleotides.

Source: GE Healthcare

RETROscript® Reverse Transcription Kit

The RETROscript® Kit provides reagents for reverse transcription of cDNA from RNA templates. When used with SuperTaq™ thermostable DNA polymerase (available separately), the cDNA templates are efficiently amplified into double-stranded DNA products. The molony-murine leukemia virus (M-MLV) reverse transcriptase is used in the first step to extend a primer hybridized to an RNA sample. M-MLV RT has higher stability and lower intrinsic RNase H activity than AMV reverse transcriptase. Both random decamers and oligo dT are provided in the RETROscript® Kit

Source: Thermo Fisher

Reverse Transcriptase M-MuLV Roche

Compared within AMV reverse transcriptase, M-MuLV reverse transcriptase lacks endonuclease activity and has much lower RNase H activity. Full-length copies of large mRNA are obtained.
M-MuLV reverse transcriptase is used for in vitro synthesis of cDNA transcripts of specific RNA sequences; for the preparation of cDNA libraries; or for synthesis of first-strand cDNA for use in subsequent amplification reactions (RT-PCR). M-MuLV reverse transcriptase is used in dideoxy-sequencing reactions in place of Klenow enzyme. Reverse transcriptase will often synthesize through GC-rich regions where Klenow enzyme is hindered.

Source: Roche

REVERSE TRANSCRIPTION SYSTEM PROMEGA

The Reverse Transcription System provides reagents to efficiently reverse transcribe RNA into cDNA in 15 minutes. The cDNA prepared from each reaction using this system may be used directly in multiple PCR amplifications using Taq DNA polymerase. The AMV Reverse Transcriptase synthesizes single-stranded cDNA from total or poly(A)+ RNA. Both Oligo(dT)15 and Random Primers are included, allowing cDNA synthesis from virtually any RNA source. The system contains sufficient reagents for 100 cDNA synthesis reactions, processing 1μg of RNA per reaction.

Source: Promega

RT2 First Strand Kit Qiagen

The RT2 First Strand Kit provides a rapid and convenient procedure for efficient first-strand cDNA synthesis and genomic DNA elimination in RNA samples. The synthesized cDNA is ready to use in real-time PCR expression analysis of multiple genes.
Qiagen also offers RT2 Easy First Strand Kits and RT2 PreAMP cDNA Synthesis Kits.

Source: Qiagen

Sensiscript Reverse Transcription Kit Qiagen

Source: Qiagen

SMART cDNA Synthesis

SMART cDNA Synthesis SMART (Switching Mechanism at 5’ End of RNA Template) is a unique technology that allows the efficient incorporation of known sequences at both ends of cDNA during first strand synthesis, without adaptor ligation. The presence of these known sequences is crucial for a number of downstream applications including amplification, RACE, and library construction. While a wide variety of technologies can be employed to take advantage of these known sequences, the simplicity and efficiency of the single-step SMART process permits unparalleled sensitivity and ensures that full-length cDNA is generated and amplified.
Source: Takara clonetech

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SuperScript® III Reverse Transcriptase

SuperScript® III Reverse Transcriptase (RT) is engineered for higher thermostability, with a longer half-life at 50°C, and reduced RNase H activity, resulting in higher yields of full-length cDNA for more complete gene product representation. If you need cDNA that represents input RNA with maximum fidelity and highest sensitivity for real time RT-PCR, you need SuperScript® III RT.
Source: Thermo Fisher

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TeloPrime Full-Length cDNA Amplification Kit

The TeloPrime Full-Length cDNA Amplification Kit is an all-in-one protocol for generating full-length cDNA from total RNA.

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ThermoScript™ RT-PCR System for First-Strand cDNA Synthesis Invitrogen

The ThermoScript™ RT-PCR System for First-Strand cDNA Synthesis Kit is optimized for generating first-strand cDNA from total or poly(A)+ RNA. ThermoScript™ Reverse Transcriptase (RT) is an avian RT with reduced RNase H activity designed for RT-PCR of the most highly structured, difficult RNA templates. The ThermoScript™ RT-PCR systems are designed to copy RNA with secondary structure and to improve specificity of priming. In conjunction with PCR, the system can be used to detect the presence of rare messages or to quantitate the amount of specific mRNA from small numbers of cells. The systems contain ThermoScript™ RT, a cloned avian reverse transcriptase that has been engineered to have reduced RNase H activity and to have high thermal stability (up to 70°C).

Source: Thermo Fisher

Transcriptor First Strand cDNA Synthesis Kit Roche

Transcriptor Reverse Transcriptase – the core component of the kit – is a recombinant reverse transcriptase expressed in E. coli. The enzyme has RNA-directed DNA polymerase activity, DNA-dependent DNA polymerase activity, unwinding activity, and RNase H activity that degrades RNA in RNA:DNA hybrids. The latter circumvents the need to perform an additional time-consuming RNase H incubation step after reverse transcription. This shortens the reaction time and reduces costs.
Transcriptor Reverse Transcriptase is recommended for RT-PCR because of its high sensitivity in combination with high thermostability: the enzyme synthesizes long cDNA products (up to 14 kb) and can be used at temperatures up to +65°C. Due to its thermostability, Transcriptor Reverse Transcriptase is recommended for GC-rich templates with high secondary structure, without the need to include additives in the reaction.

Source: Roche

Transcriptor High Fidelity cDNA Synthesis Kit Roche

The core component of the kit is the Transcriptor High Fidelity Reverse Transcriptase, a blend of a recombinant reverse transcriptase and a proofreading mediating enzyme. The synergy between both enzymes is the key to the ability of the enzyme blend to reverse transcribe RNA templates with 7-fold higher fidelity compared to other commonly used reverse transcriptases.
The Transcriptor High Fidelity Reverse Transcriptase enzyme blend efficiently reverse transcribes templates up to 14 kb. Due to the high thermostability of both enzyme components and the specially optimized buffer system, reverse transcription is possible at temperatures up to +55°C. This allows the reverse transcription of GC-rich templates with high secondary structure, without the need to include additives that may negatively influence the accuracy of the reverse transcription reaction.

Source: Roche

Universal RiboClone® cDNA Synthesis System Promega

The Universal RiboClone® cDNA Synthesis System contains the reagents required for synthesis of double-stranded cDNA from mRNA and subsequent ligation into a suitable vector. The system is based on the method described by Okayama and Berg with modifications by Gubler and Hoffman. First-strand synthesis is driven by AMV (Avian Myeloblastosis Virus) Reverse Transcriptase and either Random Primers or Oligo(dT)15 Primer, followed directly by second-strand replacement synthesis using RNase H and DNA Polymerase I.

Source: Promega


Second Strand Synthesis

NEBNext Second Strand Synthesis Module

The NEBNext Second Strand Synthesis Module has been optimized to generate double stranded cDNA from first strand cDNA. The NEBNext Second Strand Synthesis Module is provided as a master mix to maximize efficiency and convenience in RNA sample preparation workflows.
Source: NEB

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