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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.

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

TaqMan™ MicroRNA Reverse Transcription Kit (Applied Biosystems™)

The Applied Biosystems® TaqMan® MicroRNA Reverse Transcription Kit provides the necessary components for 200 reactions of optimal performance in TaqMan® MicroRNA Assays. Components of this kit are used with the RT primer provided with the TaqMan® MicroRNA Assay to convert miRNA to cDNA.

A highly specific kit that quantitates only mature miRNAs, not precursors. A sensitive kit that conserves limited samples and requires only 1 to 10 nanograms of total RNA. It is fast, simple and scalable 2-step quantitative RT-PCR assay that provides high-quality results in less than three hours.

TaqMan™ Advanced miRNA cDNA Synthesis Kit (Applied Biosystems™)

The Applied Biosystems™ TaqMan™ Advanced miRNA cDNA Synthesis Kit uses a universal reverse transcription (RT) chemistry to prepare the cDNA template for use with TaqMan™ Advanced miRNA Assays for detection and quantification of mature miRNAs in biological samples.

Features of the TaqMan Advanced miRNA workflow include:

Sensitivity: high sensitivity allows for quantification of low-expressing miRNA targets, especially in biological fluids such as plasma, serum, and tissue
Flexibility: the universal RT chemistry generates universal cDNA templates for all miRNAs present in the sample, allowing for flexibility in study design
Sample savings: requires only 2 μL of plasma/serum sample input and generates enough cDNA for 600 qPCR reactions (20 μL) or 1200 qPCR reactions (10 μL)
Specificity: 5’ ligation step allows for single base pair discrimination along entire miRNA sequence, including difficult-to-detect mismatches on 5’ end
Gold-standard TaqMan chemistry: This kit is designed for use with TaqMan® Advanced miRNA Assays for qPCR, which draw on Thermo Fisher Scientific's industry-leading bioinformatics assay design pipeline and validation system to help ensure high specificity and minimal cross-reactivity, even for closely-related miRNA families.

The TaqMan Advanced miRNA cDNA Synthesis Kit uses 3' poly-A tailing and 5' ligation of an adaptor sequence to extend the mature miRNAs present in the sample on each end prior to reverse transcription. Universal RT primers recognize the universal sequences present on both the 5' and 3' extended ends of the mature miRNAs. All mature miRNAs in the sample are reverse transcribed to cDNA.

In order to improve detection of low-expressing miRNA targets, the cDNA is then amplified using the Universal miR-Amp Primers and miR-Amp Master Mix to uniformly increase the amount of cDNA for each target, maintaining the relative differential expression levels. Unlike traditional pre-amplification, the Universal miR-Amp Primers recognize the universal sequences added to all mature miRNAs on the 5' and 3' ends, ensuring that there is no amplification bias.

Note: the TaqMan Advanced miRNA cDNA Synthesis Kit is not compatible with first generation TaqMan MicroRNA Assays for qPCR. For reverse transcription of miRNA targets using miRNA-specific RT primers, please see the TaqMan™ MicroRNA Reverse Transcription Kit.

Verso cDNA Synthesis Kit (Thermo Scientific™)

Thermo Scientific Verso cDNA synthesis Kit provides robust transcription of RNA to create a complete cDNA pool. The Verso system achieves robust and sensitive reverse transcription through the combination of a high affinity RT enzyme, a unique RNA priming method, and an optimized buffering system. The Verso RT enzyme has high RNA template affinity and reduced RNase H activity, to transcribe even sections with high secondary structure. The included anchored oligo dT priming method further enhances sensitivity by increasing transcription efficiency, and the cDNA synthesis buffer has been optimized to achieve a full and diverse cDNA pool.

Highlights

• Highly sensitive with a broad dynamic range: can reverse transcribe RNA from an input range of 1pg to 1µg
• Efficient RT reaction allows for 75% shorter protocol times (down to 30 minutes)
• Unique priming strategy for increased efficiency and broad template flexibility
• High affinity RT enzyme effectively transcribes through difficult RNA sequences
• Wide working temperature range (42°C to 57°C) for success with GC-rich and other difficult templates
• Included RT Enhancer prevents genomic DNA carryover, eliminating the need for separate DNase I treatment
• RNase Inhibitor included
• Can be paired with any PCR Kit for complete 2-Step RT-(q)PCR

Applications

• 2-step RT-(q)PCR
• Single stranded cDNA synthesis

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Verso cDNA Synthesis Kit

RevertAid RT Reverse Transcription Kit (Thermo Scientific™)

Thermo Scientific RevertAid RT Kit is a complete system for efficient synthesis of first strand cDNA from mRNA or total RNA templates. The kit uses RevertAid Reverse Transcriptase (RT), which has lower RNase H activity compared to AMV reverse transcriptase. The enzyme maintains activity at 42-50°C and is suitable for synthesis of cDNA up to 13 kb. The recombinant Thermo Scientific RiboLock RNase Inhibitor, supplied with the kit, effectively protects RNA from degradation at temperatures up to 55°C. First strand cDNA synthesized with this system can be directly used as a template in PCR or real-time PCR. It is also ideal as a first step for second strand cDNA synthesis or linear RNA amplification. Radioactively and non-radioactively labeled nucleotides can be incorporated into first strand cDNA for use as a probe in hybridization experiments, including microarrays.

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™ First-Strand Synthesis System for RT-PCR (Invitrogen™)

The SuperScript® First-Strand Synthesis System for RT-PCR is optimized to synthesize first-strand cDNA from purified poly(A)+ or total RNA. The system can be used with as little as 1 ng or as much as 5 µg of total RNA. After synthesis, the cDNA can be amplified with specific primers by PCR without intermediate organic extractions or ethanol precipitations. In conjunction with PCR, the system can be used to detect the presence of rare messages, to quantitate the amount of specific mRNA from small numbers of cells, or to clone specific cDNAs without constructing an entire cDNA library. The system is flexible, allowing the use of any PCR enzyme. Combine with AccuPrime™ Taq DNA Polymerase or Platinum® Taq DNA Polymerase for higher specificity PCR or with AccuPrime™ Pfx DNA Polymerase for high-fidelity cloning applications.

SuperScript II RT
The first-strand cDNA synthesis reaction is catalyzed by SuperScript® II Reverse Transcriptase (RT). This enzyme has been engineered to reduce the RNase H activity that degrades mRNA during the first-strand reaction, resulting in greater full-length cDNA synthesis and higher yields of first-strand cDNA than obtained with RNase H+ RTs. Because SuperScript® II RT is not inhibited significantly by ribosomal and transfer RNA, it may be used effectively to synthesize first-strand cDNA from a total RNA preparation. The enzyme exhibits increased thermal stability and may be used at temperatures up to 50°C.

Using the SuperScript® First-Strand Synthesis System for RT-PCR
This system has been optimized to synthesize first-strand cDNA from varying amounts of starting material. The SuperScript® II RT concentration has been lowered and RNaseOUT™ Recombinant RNase Inhibitor has been added to the system as part of this optimization process. Additionally, reaction conditions have been modified to further increase the sensitivity of the system. Using the kit, you synthesize first-strand cDNA using either total RNA or poly(A)+-selected RNA primed with oligo(dT), random primers, or a gene-specific primer. Then, you perform PCR in a separate tube using primers specific for the gene of interest.

SuperScript™ III First-Strand Synthesis SuperMix (Invitrogen™)

The SuperScript® III First-Strand Synthesis SuperMix is an optimized SuperMix formulation for first-strand cDNA synthesis from purified poly(A)+ or total RNA. RNA targets from 100 bp to >12 kb can be detected with this system. The amount of starting material can vary from 0.1 pg to 5 µg of total RNA.

Using the SuperScript® III First-Strand Synthesis SuperMix
The kit includes SuperScript® III/RNaseOUT™ Enzyme Mix, 2X First-Strand Reaction Mix, and Annealing Buffer. SuperScript® III Reverse Transcriptase, included in the Enzyme Mix, is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme can be used to synthesize cDNA at a temperature range of 45–55°C, providing increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. Because SuperScript® III RT is not significantly inhibited by ribosomal and transfer RNA, it can be used to synthesize cDNA from total RNA. RNaseOUT™ Recombinant RNase Inhibitor is included in the Enzyme Mix to safeguard against degradation of target RNA due to ribonuclease contamination. The 2X First-Strand Reaction Mix includes 10 mM MgCl2 and 1 mM of each dNTP in a buffer formulation that has been optimized for first-strand synthesis of cDNA. The Annealing Buffer is used in the initial template-primer annealing step. Separate tubes of oligo(dT)20 and random hexamers are also provided. cDNA synthesis can be performed using either total RNA or poly(A)+-selected RNA primed with oligo(dT), random primers, or a gene-specific primer.

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

The High Capacity cDNA Reverse Transcription Kit 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. 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.

SuperScript™ III First-Strand Synthesis System (Invitrogen™)

The SuperScript® III First-Strand Synthesis System for RT-PCR is optimized to synthesize first-strand cDNA from purified poly(A)+ or total RNA. RNA targets from 100 bp to >12 kb can be detected with this system. The amount of starting material can vary from 1 pg–5 µg of total RNA. SuperScript® III Reverse Transcriptase is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme is used to synthesize cDNA at a temperature range of 42–55°C, providing increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. Because SuperScript® III RT is not significantly inhibited by ribosomal and transfer RNA, it may be used to synthesize first-strand cDNA from a total RNA preparation.

Using the SuperScript® III First-Strand System
cDNA synthesis is performed in the first step using either total RNA or poly(A)+-selected RNA primed with oligo(dT), random primers, or a gene-specific primer. In the second step, PCR is performed in a separate tube using primers specific for the gene of interest. For the PCR reaction, we recommend one of the following DNA polymerases: Platinum® Taq DNA Polymerase provides automatic hot-start conditions for increased specificity up to 4 kb, Platinum® Taq DNA Polymerase High Fidelity provides increased yield and high fidelity for targets up to 15 kb, and Platinum® Pfx DNA Polymerase provides maximum fidelity for targets up to 12 kb.

SuperScript™ IV First-Strand Synthesis System (Invitrogen™)

The SuperScript® IV First-Strand Synthesis System for RT-PCR is optimized for synthesis of first-strand cDNA from purified poly(A)+ or total RNA. The kit includes SuperScript® IV Reverse Transcriptase (RT), a proprietary MMLV mutant that produces robust and reliable cDNA synthesis in RT reactions. The SuperScript® IV synthesis system is significantly improved over the SuperScript® III-based system in inhibitor resistance, processivity, and reaction speed, while retaining all the benefits of the previous system, including increased thermostability, highly efficient full-length cDNA synthesis, and reduced RNase activity. The SuperScript® IV system is designed to provide reliable, consistent, and fast cDNA synthesis in the presence of inhibitors found in a wide variety of samples that cause other currently available RTs to perform inefficiently.

The SuperScript® IV First-Strand Synthesis System contains all components needed for RT reactions, plus an additional control gene and primers, and provides the flexibility to customize the RT set-up to fit the needs of your application. The SuperScript® IV synthesis system is the top choice for performance and flexibility for RT-PCR applications. We recommend it for all RT-PCR applications, especially when reproducibility and reliability is the primary concern and when inhibitors in the RNA sample may interfere with cDNA synthesis, leading to biases in gene expression studies.

Features of the SuperScript® IV First-Strand Synthesis System include:
• Significantly improved resistance to a variety of inhibitors that can interfere with cDNA synthesis
• Robust and specific cDNA synthesis in a wide range of sample types
• A faster reverse transcriptase reaction that reduces incubation time from >50 min to 10 min
• Significantly better processivity compared to SuperScript® III RT

Enzyme source
Purified from E. coli expressing the pol gene of M-MLV, modified to increase thermostability and half-life, processivity, inhibitor resistance and to reduce RNase H activity.

Performance and quality testing
Endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product.

Unit definition
One unit of SuperScript® IV RT is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min at 37°C using poly(A) oligo(dT)12-18 as a template/primer.

Unit reaction conditions
50 mM Tris-HCl (pH 8.3), 4 mM MgCl2, 10 mM DTT, 50 mM KCl, 0.5 mM dTTP, 0.4 MBq/mL [3H]-dTTP, 0.4 mM poly(A) oligo (dT)12-18 and enzyme in 20 μL for 10 min at 37°C.

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.

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.

SuperScript™ III First-Strand Synthesis SuperMix for qRT-PCR (Invitrogen™)

SuperScript® III First-Strand Synthesis SuperMix for qRT-PCR provides the high-temperature capability of SuperScript® III Reverse Transcriptase in an optimized SuperMix format for the synthesis of first-strand cDNA for use in real-time quantitative RT-PCR (qRT-PCR). The simple, time-saving reaction set-up uses just two tubes: a 2X Reaction Mix and an Enzyme Mix.

Enzyme mix
SuperScript® III Reverse Transcriptase, included in the RT Enzyme Mix, is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme can be used to synthesize cDNA at a temperature range of 42–60°C, providing increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. Because SuperScript® III RT is not significantly inhibited by ribosomal and transfer RNA, it can be used to synthesize cDNA from total RNA. RNaseOUT™ Recombinant Ribonuclease Inhibitor, also included in the enzyme mix, is an RNase inhibitor protein that safeguards against the degradation of target RNA due to ribonuclease contamination of the RNA preparation.

Reaction mix
The 2X RT Reaction Mix includes oligo(dT)20, random hexamers, MgCl2, and dNTPs in a buffer formulation that has been optimized for qRT-PCR. E. coli RNase H is provided as a separate tube in the kit to remove the RNA template from the cDNA:RNA hybrid molecule after first-strand synthesis. This has been shown to increase sensitivity in qRT-PCR.

Using SuperScript® III First-Strand Synthesis SuperMix
This SuperMix formulation can be used to quantify fewer than 10 copies of a target gene in qRT-PCR, with a broad dynamic range that supports accurate quantification of high-copy mRNA from up to 1 µg of total RNA. Reagents are provided for 50 or 250 RT reactions of 20 µL each.

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.

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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™ III CellsDirect™ cDNA Synthesis Kit (Invitrogen™)

The SuperScript® III CellsDirect cDNA Synthesis Kit is optimized for synthesizing first-strand cDNA directly from a mammalian cell lysate without first isolating the RNA. Lysis and reverse transcription are performed in the same tube, and the resulting first-strand cDNA is ready to use in cloning and PCR. For real-time quantitative RT-PCR, see the note below.

Advantages of the SuperScript® III CellsDirect cDNA Synthesis Kit include:

• Compatible with a wide range of mammalian cell types grown under different treatment conditions
• Single-tube format minimizes reagent loss, sample loss, and handling time
• Total lysate volume is used in first-strand cDNA synthesis reaction, providing greater yields with a limited number of cells and allowing for detection of rare transcripts
• SuperScript® III Reverse Transcriptase, with reduced RNase H activity and higher thermal stability, produces high yields of cDNA in the first-strand synthesis reaction, for greater sensitivity and enhanced detection of rare transcripts
• Generates high-quality cDNA for use in a variety of applications, including cloning and PCR
• Simple protocol takes less than 2 hours

How it works
In traditional RT-PCR, RNA is first isolated from cells in a time-consuming procedure that can lead to a loss of material. Using the SuperScript® III CellsDirect cDNA Synthesis System, the cells are lysed and the cDNA is generated from the lysate in a single tube with minimal handling and no sample loss. DNase I is added to eliminate genomic DNA prior to first-strand synthesis. This kit has been optimized for small cell samples, ranging from 10,000 cells down to a single cell (as measured by serial dilution). The use of SuperScript® III Reverse Transcriptase ensures high specificity and high yields of cDNA from small amounts of starting material—as little as 10 pg total RNA. After synthesis, the first-strand cDNA can be amplified with specific primers by PCR without intermediate organic extractions or ethanol precipitations.