Shop All cDNA Synthesis Kits

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™ 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 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

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.

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.

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.

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).

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.

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.

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. The kit has the following advantages:

• 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.

TaqMan™ MicroRNA Reverse Transcription Kit Applied Biosystems™

The Applied Biosystems® TaqMan® MicroRNA Reverse Transcription Kit provides the necessary components for 1000 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.

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.

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.

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™ 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.
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