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Maxima™ H Minus cDNA Synthesis Master Mix (Thermo Scientific™)

Thermo Scientific Maxima H Minus cDNA Synthesis Master Mix provides all cDNA synthesis reaction components in a convenient one-tube master mix. It is optimized for highly efficient cDNA synthesis for two-step quantitative RT-PCR (RT-qPCR) applications.

Features include:
• One-tube master mix helps reduce pipetting steps and enhanced consistency in RT-qPCR results
• Increased RT efficiency across a wide range of input RNA amounts and gene targets
• High thermostability to allow RT reactions at 50 to 65°C temperature range

The master mix contains Maxima H Minus Reverse Transcriptase (RT) and RiboLock RNase Inhibitor. Maxima H Minus RT is an advanced enzyme derived from M-MuLV RT by in vitro evolution. The enzyme features high thermostability, robustness, processivity, increased cDNA synthesis rate, and lack of RNase H activity. The recombinant RiboLock RNase Inhibitor effectively protects RNA template from degradation by RNases A, B, and C at temperatures up to 55°C. The master mix also contains reaction buffer, dNTPs, oligo (dT)18, and random hexamer primers. A separate tube of No-RT control mix is also provided.

Maxima H Minus cDNA Synthesis Master Mix is capable of reproducible cDNA synthesis at elevated temperatures (50–65°C). The synthesis reaction is typically complete in 15–30 minutes.

Additional information about reaction components
• The No RT Control mix contains all the components in the Maxima H Minus cDNA Synthesis Master Mix except RT. The presence of a PCR product in the No RT Control reaction indicates that the reaction is contaminated with DNA. To further enhance genomic DNA elimination efficiency, template RNA incubation with dsDNase (Cat. No.EN0771) is strongly recommended.
• Nuclease-free water is provided for reaction set-up and dilution of sample DNA. The absence of exodeoxyribonucleases, ribonucleases, and phosphatases has been confirmed by appropriate quality tests.

Related products
Maxima H Minus First Strand cDNA Synthesis Kit, with dsDNase
Maxima H Minus First Strand cDNA Synthesis Kit
Maxima H Minus Reverse Transcriptase
dsDNase

SuperScript™ VILO™ cDNA Synthesis Kit (Invitrogen™)

The SuperScript VILO cDNA Synthesis Kit is designed to generate first strand cDNA for two-step RT-qPCR applications. The kit is supplied in a 2-tube format with the VILO Reaction Mix and an enzyme blend in separate tubes. The enzyme blend contains SuperScript III Reverse Transcriptase (RT), a genetically engineered MMLV RT that has reduced RNase H activity and improved thermostability for highly efficient cDNA synthesis. The kit can be used to synthesize cDNA from for a wide range of input RNA amounts.

Note: For superior first-strand cDNA synthesis performance in two-step RT-qPCR, we recommend SuperScript IV VILO Master Mix or SuperScript IV VILO Master Mix with ezDNase Enzyme. The master mix provides all cDNA synthesis reaction components in a convenient one-tube master mix format. It elevates the trusted VILO technology to the next level with the highly processive and thermostable SuperScript IV RT and further optimized buffer. The master mix enables efficient cDNA synthesis at higher temperatures in less time and provides superior cDNA yield and sensitivity even with suboptimal purity or scarce templates.

Maxima First Strand cDNA Synthesis Kit Reaction Mix (5X) (Thermo Scientific™)

Thermo Scientific Maxima 5X Reaction Mix is a component of the Maxima First Strand cDNA Synthesis Kits for RT-qPCR (K1671/K1672/K1641/K1642) and may be purchased separately. The 5X Reaction Mix is comprised of reaction buffer, dNTPs, oligo (dT)18 and random hexamer primers.

Related Products
Maxima First Strand cDNA Synthesis Kit for RT-qPCR, with dsDNase
Maxima First Strand cDNA Synthesis Kit for RT-qPCR

RevertAid First Strand cDNA Synthesis Kit (Thermo Scientific™)

Thermo Scientific RevertAid First Strand cDNA Synthesis Kit is a complete system for efficient synthesis of first strand cDNA from RNA templates. The kit uses RevertAid Reverse Transcriptase (RT), a recombinant M-MuLV RT which maintains activity at 42-50°C and is suitable for synthesis of cDNA up to 13 kb. RiboLock RNase Inhibitor, supplied with the kit, effectively protects RNA templates from degradation.
The RevertAid First Strand cDNA Synthesis Kit is supplied with both oligo(dT)18 and random hexamer primers. The oligo(dT)18 primer anneals selectively to the poly(A) tail of mRNA. Random hexamer primers do not require the presence of the poly(A) tail, therefore, they can be used for transcription of the 5'-end regions of mRNA or cDNA synthesis of RNA species lacking a poly(A) tail (e.g., microRNAs). Gene-specific primers may also be used with the kit.

Highlights

• Full-length first strand cDNA up to 13 kb
• Optimum reaction temperature 42°C
• Complete kit—all the components for the RT reaction are included

Applications

• First strand cDNA synthesis for RT-PCR and RT-qPCR
• Construction of full length cDNA libraries
• Antisense RNA synthesis

SuperScript™ Choice System for cDNA Synthesis (Invitrogen™)

The SuperScript® Choice System is designed for synthesis of double-stranded cDNA from a purified mRNA population. Advantages of the SuperScript® Choice system include:

• The advanced capabilities of SuperScript® II RT resulting in greater cDNA yields and increased full-length cDNAs (1,3)
• Choice of primers for first-strand synthesis (2)
• One-tube format improving cDNA yield (4,5)
• cDNA is size-fractionated using an efficient and convenient pre-packed column
• One reaction converts up to 5 µg of mRNA into size-fractionated, EcoR I-adapted cDNA, ready for ligation into any EcoR I-digested vector

Performance and Quality Testing: Yields of the first- and secondstrand cDNA synthesis reactions and percentage of full-length double-stranded cDNA are assessed using the control RNA.

SuperScript™ Double-Stranded cDNA Synthesis Kit (Invitrogen™)

The SuperScript® Double-Stranded cDNA Synthesis Kit contains all of the reagents (except an oligo (dT)-containing primer) necessary to make high-quality, double-stranded cDNA from total RNA or poly A+-selected RNA (mRNA). The advanced capabilities of SuperScript® II Reverse Transcriptase, with its reduced RNase H activity, maximize the yield of full-length cDNA, as well as that of overall cDNA synthesis. Features of the SuperScript® Double-Stranded cDNA Synthesis Kit include:

Reliability—each buffer, reagent, and enzyme in the kit is of the same high quality you expect from us
Performance—SuperScript® II is an enzyme you can trust for high-yield, high-quality, general-purpose, first-strand synthesis.

Simple, reliable, and trouble-free
The SuperScript® Double-Stranded cDNA Synthesis Kit offers a simple, reliable, and trouble-free method by which RNA templates can be converted to cDNA for use in cloning and library construction experiments. The reactions described in the SuperScript® Double-Stranded cDNA Synthesis Kit protocol are designed to convert 25–50 µg total RNA, or 0.2–5 µg mRNA, into first- and second-strand cDNA (supply your own primer). Enjoy the benefits of a tested protocol and reagents of consistent quality. To get the best possible cDNA product using this kit, we recommend the use of a high-quality RNA substrate prepared using TRIzol® RNA Isolation Reagents.

Maxima™ H Minus cDNA Synthesis Master Mix, with dsDNase (Thermo Scientific™)

Thermo Scientific Maxima H Minus cDNA Synthesis Master Mix with dsDNase provides a simple workflow that combines genomic DNA elimination and cDNA synthesis in a one-tube procedure. The cDNA reaction components are pre-mixed into a complete master mix that is convenient to use, reduces pipeting steps, and is optimized for cDNA synthesis in two-step quantitative RT-PCR (RT-qPCR) applications. Maxima H Minus cDNA Synthesis Master Mix is optimized for reproducible cDNA synthesis at elevated temperatures (50–65°C) within 30 minutes.

Features include:
• One-tube master mix for reduced pipetting steps and enhanced consistency in RT-qPCR results
• Increased RT efficiency across a wide range of input RNA amounts and gene targets
• High thermostability to allow RT reactions at 50 to 65°C temperature range
• Integrated step of genomic DNA removal from RNA samples

The master mix contains Maxima H Minus Reverse Transcriptase (RT) and RiboLock RNase Inhibitor. Maxima H Minus RT is an advanced enzyme derived from M-MuLV RT by in vitro evolution. The enzyme features high thermostability, robustness, processivity, increased cDNA synthesis rate, and lack of RNase H activity. The recombinant RiboLock RNase Inhibitor effectively protects RNA template from degradation by RNases A, B, and C at temperatures up to 55°C. The master mix also contains reaction buffer, dNTPs, oligo (dT)18, and random hexamer primers. A separate tube of No-RT control mix is provided for convenient RT negative control.

The included double-strand specific DNase (dsDNase) allows removal of genomic DNA from RNA samples in 2 minutes without affecting the quality or quantity of RNA. This dsDNase is also available separately (Cat. No. EN0771).

Additional information about reaction components
• The No RT Control mix contains all components in the Maxima H Minus cDNA Synthesis Master Mix except Maxima H Minus RT. The presence of a PCR product in the No RT Control reaction indicates that the reaction is contaminated with DNA. To further enhance genomic DNA elimination efficiency, template RNA incubation with the included dsDNase is strongly recommended.
• The dsDNase is used for rapid and safe removal of contaminating genomic DNA from RNA samples. dsDNase is easily inactivated by moderate heat treatment (55°C). It allows for a dramatically simplified workflow that combines genomic DNA elimination and cDNA synthesis into one-tube procedure.
• Nuclease-free water is provided for reaction set-up and dilution of sample DNA. The absence of exodeoxyribonucleases, ribonucleases, and phosphatases has been confirmed by appropriate quality tests.

Related products
Maxima H Minus First Strand cDNA Synthesis Kit, with dsDNase
Maxima H Minus First Strand cDNA Synthesis Kit
Maxima H Minus Reverse Transcriptase
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RevertAid H Minus First Strand cDNA Synthesis Kit (Thermo Scientific™)

Thermo Scientific RevertAid H Minus First Strand cDNA Synthesis Kit is a complete system for efficient synthesis of first strand cDNA from RNA templates. The kit includes RevertAid H Minus Reverse Transcriptase, which has a point mutation that completely eliminates RNase H activity. Therefore, it does not degrade RNA in RNA-DNA hybrids during synthesis of the first strand cDNA.

Highlights

• High yields of full-length first strand cDNA up to 13 kb
• Increased reaction temperatures in the range of 42 to 55°C
• Supplied with the recombinant RiboLock RNase Inhibitor
• Complete kit—oligo(dT)18 and random hexamer primers included with the kit

Applications

• First strand cDNA synthesis for RT-PCR and real-time RT-PCR
• Construction of full length cDNA libraries
• Antisense RNA synthesis

Additional Features

The recombinant RiboLock RNase Inhibitor, supplied with the kit, effectively protects RNA template from degradation. It is fully compatible with the reverse transcription reaction, as it maintains activity at temperatures up to 55°C.The kit is supplied with both oligo(dT)18 and random hexamer primers. The oligo(dT)18 primer anneals selectively to the poly(A) tail of mRNA. Random hexamer primers do not require the presence of poly(A), therefore, they can be used for transcription of the 5’-end regions of mRNA or for cDNA synthesis using RNA without the poly(A) tail, e.g., microRNAs. Gene-specific primers may also be used with the kit. The first strand cDNA can be directly used as a template in PCR, realtime PCR or in second strand cDNA synthesis.

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.

Features of this kit include:
Fast—short reaction time (typically 30–60 min)
Convenient—easy workflow with few pipetting steps (2 tubes)
Reliable—reverse transcription of both abundant and limited targets
Flexible—optimized 2-step protocol, enabling multiple PCR reactions from a single reverse transcription reaction

Streamline your cDNA synthesis while maintaining detection sensitivity
The components of the two-tube kit work together to provide sensitive and specific reverse transcription across a broad range of template amounts. Amplification of a dilution series of the RNA concentration reference standard demonstrates exceptional reverse transcription and PCR efficiency across 11 orders of magnitude. This wide dynamic range allows one set of reaction conditions to detect transcripts from highly active as well as weakly expressed genes. The two-tube formulation reduces reaction time and requires fewer pipetting steps than with the High Capacity cDNA Reverse Transcription Kit, while maintaining high performance.

Arcturus™ RiboAmp™ HS PLUS cDNA Kit (Applied Biosystems™)

The ARCTURUS® RiboAmp® HS PLUS cDNA Kit permits quantitative Real-Time PCR (qRT-PCR) from as little as 100pg of total RNA. The kit uses random primers to enable robust reverse transcription across a broader transcript length.

Integrated Systems for Microgenomics
The ARCTURUS® Complete System for Microgenomics® offers an integrated solution for preparing small quantities of RNA for gene expression analysis. The System features the ARCTURUSXT™ Laser Capture Microdissection (LCM) Instrument to procure pure cell populations, the PicoPure® RNA Isolation Kit to extract and purify RNA, and the RiboAmp® PLUS Kit for reverse transcription and linear amplification of RNA. ARCTURUS® Systems for Microgenomics enable accurate and sensitive qRT-PCR and microarray assays that reveal molecular otherwise obscured in whole tissue samples or cell mixtures.

For Research Use Only. Not for use in diagnostics procedures.

High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor (Applied Biosystems™)

The High Capacity cDNA Reverse Transcription Kit with RNase Inhibitor contains all components necessary for the quantitative conversion of up to 2 µg of total RNA to single-stranded cDNA in a single 20 µL reaction. This particular kit also includes 5 tubes of RNase inhibitor (200 µL each).

The High Capacity cDNA Reverse Transcription Kit delivers extremely high-quality, single-stranded cDNA from 0.02 to 2 µg total RNA. Reactions can be scaled up to 100 µL to generate 10 µg of cDNA from a single reaction. Downstream applications include real-time PCR, standard PCR, and microarrays. The kit is ideal for generating cDNA archives.

Features of this kit include:

• Linear target amplification for real-time PCR
• Higher yields and precision than other cDNA synthesis kits at a fraction of the cost
• 10-fold greater dynamic range than other kits

Extensively tested with a variety of templates
Quantitative first-strand synthesis of all RNA species is achieved with the use of the random primers. The kit has been extensively tested with a variety of RNA templates, including GC- and AU-rich targets and RNAs expressed at low levels. cRNA can also be efficiently generated from in vitro transcription of cDNA.

Cells-to-cDNA™ II Kit (Invitrogen™)

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 100 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.

• No RNA purification required
• From cells in culture to cDNA in less than 2 hr
• Detect rare messages in as few as 1 cell
• Ideal for labs not equipped for RNA isolation

Ideal for RT-PCR Applications
The Cells-to-cDNA™ II Kit (patent pending) integrates RNase inactivation and DNase I treatment into an RT-PCR compatible cell lysis buffer, eliminating RNA isolation altogether. Most cell lysis buffers contain strong denaturants that if carried over into enzymatic reactions would inhibit or inactivate most enzymes. If a lysis buffer without strong denaturants is used, endogenous RNase can quickly degrade cellular RNA. The Cells-to-cDNA™ II Kit contains a novel Cell Lysis Buffer that inactivates endogenous RNases without compromising downstream enzymatic reactions. After inactivation of RNases, the cell lysate can be directly added to a cDNA synthesis reaction. Cells-to-cDNA™ II is compatible with both one-step and two-step RT-PCR protocols.

Quantitative—Linear Results and No Detectable DNA Contamination
To illustrate the quantitative, linear response of Cells-to-cDNA™ II to variations in input cell number, a real-time quantitative RT-PCR experiment was performed using the ABI 7700 Sequence Detection System. Plotting cycle threshold (Ct) versus cell concentration for GAPDH yielded a standard curve with a correlation of 0.99. Thus, Cells-to-cDNA™ II yields linear results for 1 to 10,000 cells/ µL in a real-time RT-PCR two-step assay. It should also be noted that the minus-template PCR control for the GAPDH experiment was negative, indicating complete removal of genomic DNA.

Simple Procedure
The kit comes with everything you need to go from cultured cells to PCR-ready cDNA in less than 2 hr; no RNA isolation is required. Cells are first washed with PBS and then resuspended in the Cell Lysis Buffer. A brief heating simultaneously lyses the cells and inactivates RNases. An optional DNase digestion is then performed to degrade genomic DNA, followed by a heat inactivation step. A fraction of the cell lysate is then used in a reverse transcription reaction with the included reagents.

Accessory Products:
SuperTaq™ Thermostable Taq DNA Polymerase is recommended and available separately (SKU#s AM2050 and AM2052). For optimal amplification of fragments greater than 1 kb, use SuperTaq™ Plus Thermostable Taq DNA Polymerase (SKU#s AM2054 and AM2056).

Maxima First Strand cDNA Synthesis Kit for RT-qPCR, with dsDNase (Thermo Scientific™)

Thermo Scientific Maxima First Strand cDNA Synthesis Kit for RT-qPCR with dsDNase is a convenient system optimized for cDNA synthesis in 2-step quantitative RT-PCR (RT-qPCR) applications. It provides a dramatically simplified workflow that combines genomic DNA elimination and cDNA synthesis into one-tube procedure. The kit contains a novel double-strand specific DNase (dsDNase) engineered to remove contaminating genomic DNA from RNA preps in 2 minutes without damage to quality or quantity of RNA. Highly specific activity towards double-stranded DNA ensures that single-stranded DNA (such as cDNA and primers) is not cleaved and dsDNase treated RNA can be directly added to reverse transcription.

For the first strand cDNA synthesis the kit uses Maxima Reverse Transcriptase (RT), an advanced enzyme derived by in vitro evolution of M-MuLV RT. The enzyme features high thermostability, robustness and increased cDNA synthesis rate compared to wild type M-MuLV RT. The Maxima First Strand cDNA Synthesis Kit for RT-qPCR is capable of reproducible cDNA synthesis from a wide range of total RNA amounts (1 pg to 5 µg) at elevated temperatures (42 to 65°C). The synthesis reaction can be completed in 15 to 30 minutes. Components of the Maxima First Strand cDNA Synthesis Kit for RT-qPCR are pre-mixed to save time and to reduce the possibility of pipetting errors.

Highlights

• Integrated gDNA removal step
• High yields of full length cDNA up to 20 kb.
• Efficient cDNA synthesis at wide temperature range from 42°C to 65°C.
• Increased synthesis rate—complete cDNA synthesis in 15-30 minutes.
• High sensitivity and specificity.

Applications

• 2-step RT-PCR
• 2-step RT-qPCR

Maxima First Strand cDNA Synthesis Kit with dsDNase contains
• Maxima Enzyme Mix
• 5X Reaction Mix
• dsDNase
• 10X dsDNase Buffer
• nuclease-free water

Additional Information about Reaction Components
The Maxima Enzyme Mix contains Maxima Reverse Transcriptase and Thermo Scientific RiboLock RNase Inhibitor,. The recombinant RiboLock RNase Inhibitor effectively protects RNA templates from degradation by RNases A, B and C at temperatures up to 55°C.

5X Reaction Mix contains the remaining reaction components: reaction buffer, dNTPs, oligo(dT)18 and random hexamer primers.

Water, nuclease-free is provided for reaction set-up and dilution of sample RNA. The absence of endo-, exodeoxyribonucleases, ribonucleases and phosphatases has been confirmed by appropriate quality tests.

Related Products
Maxima First Strand cDNA Synthesis Kit for RT-qPCR, with dsDNase
Maxima First Strand cDNA Synthesis Kit for RT-qPCR
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TaqMan™ Reverse Transcription Reagents (Invitrogen™)

TaqMan® Reverse Transcription Reagents provide the necessary reagents for reverse transcriptase-PCR (RT-PCR). Random hexamers, oligo (dT)16, and reverse primers are included for cDNA synthesis flexibility. Advantages of TaqMan® Reverse Transcription Reagents:

• Contain MultiScribe® Reverse Transcriptase, a recombinant moloney murine leukemia virus reverse transcriptase (MMLV RT)
• Random hexamers and oligo (dT)16 included for flexibility
• Individual components provide versatility in assay set-up
• Compatible with two-step RT-PCR reactions

Using TaqMan® Reverse Transcription Reagents
TaqMan® Reverse Transcription Reagents provide all the necessary components to perform the reverse transcription of RNA to cDNA. For two-step RT-PCR, random hexamers, oligo (dT)16, and sequence-specific reverse primers can all be used. The choice of primers for reverse transcription is best made after experimentally evaluating all three priming systems.

Maxima H Minus First Strand cDNA Synthesis Kit, with dsDNase (Thermo Scientific™)

Thermo Scientific Maxima H Minus First Strand cDNA Synthesis Kit with dsDNase provides a dramatically simplified workflow that combines genomic DNA elimination and cDNA synthesis into one-tube procedure. The kit contains a novel double-strand specific DNase (dsDNase) engineered to remove contaminating genomic DNA from RNA preps in 2 minutes without damage to quality or quantity of RNA. Highly specific dsDNase activity towards double-stranded DNA ensures that single-stranded DNA (such as cDNA and primers) is not cleaved and dsDNase treated RNA can be directly added to reverse transcription.

For reverse transcription the kit uses Thermo Scientific Maxima H Minus Reverse Transcriptase (RT), which is an advanced enzyme derived by in vitro evolution of M-MuLV RT. The enzyme features the highest thermostability among the derivatives of M-MuLV RT and lacks RNase H activity. The Maxima H Minus First Strand cDNA Synthesis Kit allows synthesis of long cDNAs up to 20 kb at elevated temperatures (up to 65°C), superseding other systems' abilities to produce full-length cDNA. Due to increased synthesis rates the reaction can be completed in 30 minutes.

Highlights

• Integrated genomic DNA removal step
• Increased reaction temperatures—the first strand of cDNA can be synthesized within the 42 to 65°C temperature range
• High yields of full-length first strand cDNA—with RNA templates up to 20 kb
• Flexible priming—oligo(dT)18, random hexamer, or gene-specific primers

Applications

• First Strand cDNA synthesis for RT-PCR
• Construction of cDNA libraries
• Generation of probes for hybridization
• Antisense RNA synthesis

Includes

Maxima H Minus First Strand cDNA Synthesis Kit with dsDNase contains Maxima H Minus Enzyme Mix, dsDNase, 10X dsDNase Buffer, Oligo(dT)18 and Random hexamer primers, 5X RT Buffer, dNTP Mix, and nuclease-free water.

Additional information about reaction components

• Maxima H Minus Enzyme Mix contains Maxima H Minus Reverse Transcriptase and RiboLock RNase Inhibitor. RiboLock RNase Inhibitor effectively protects RNA templates from degradation by RNases A, B and C at temperatures up to 55°C.
oligo(dT)18 and random hexamer primers are supplied with the kit. Random hexamer primers bind non-specifically and are used to synthesize cDNA from all RNAs in a total RNA population. The oligo(dT)18 primer selectively anneals to the 3'-end of poly(A) RNA, synthesizing cDNA only from poly(A) tailed mRNA. Gene-specific primers may also be used with the kit to prime synthesis from a specified sequence.
10 mM dNTP Mix is a premixed aqueous solution of dATP, dTTP, dCTP, and dGTP.
Nuclease-free water is provided for reaction set-up and dilution of sample DNA. The absence of endo-, exodeoxyribonucleases, ribonucleases, and phosphatases has been confirmed by appropriate quality tests.

Related Products
Maxima H Minus First Strand cDNA Synthesis Kit, with dsDNase
Maxima H Minus First Strand cDNA Synthesis Kit