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Reverse Transcription |
Reverse transcription is the process of transcribing RNA molecules into complementary DNA. This cDNA synthesis reaction is mediated by enzymes called reverse transcriptases, or RTs. Moloney murine leukemia virus (M-MLV) reverse transcriptase is one of the RTs used in molecular biology workflows. However, novel engineered RTs are often preferred for enhanced efficiency, improved thermostability, and higher cDNA yields.
How do reverse transcriptases work?
Reverse transcription reactions involve multiple steps to synthesize cDNA. Reverse transcriptases exhibit several distinct biochemical activities, predominantly RNA-dependent DNA polymerase activity and RNase H activity. Firstly, reverse transcriptase binds to an RNA template in the presence of an annealed primer. Next, the RNA-dependent DNA polymerase activity of the enzyme synthesizes cDNA by incorporating dNTPs from the reaction mix. This generates a cDNA:RNA hybrid and is referred to as first-strand cDNA synthesis. If reverse transcriptase exhibits RNase H activity (as in wild-type M-MLV RT), the RNA of the cDNA:RNA hybrid is cleaved during first-strand synthesis. In engineered enzymes, RNase H activity is often reduced to prevent premature RNA degradation.
RT enzymes and cDNA synthesis kits
Choose from a variety of reverse transcriptase stand-alone enzymes, master mixes, cDNA synthesis kits, primers, and dNTP solutions to suit your application. Invitrogen SuperScript IV products aid in exceptional cDNA synthesis from any type of template RNA.
Invitrogen SuperScript product family is ideal for all cDNA synthesis experiments. SuperScript IV RT offers top-notch performance marked by thermostability, high processivity, and exceptional cDNA yields, and is available in a selection of formats.
First strand cDNA synthesis master mix crafted with VILO (Variable Input, Linear Output) technology is an ideal choice for two-step RT-qPCR applications. This master mix is known for its remarkable sensitivity, exceptional linearity, and short reaction times.
Learn more about cDNA synthesis master mix for two-step RT-qPCR
One-step RT-PCR kits are optimized for cDNA synthesis and amplification in a single step. This approach is often preferred for a faster and simpler workflow and is suitable for high throughput applications.
Skip RNA isolation and expedite cDNA synthesis. Direct reverse transcription products allow cell lysis and cDNA synthesis to be performed in a single tube, using as few as single cell samples.
RNase inhibitors inhibit RNases that degrade template RNA. Use RNase inhibitors to maintain RNA integrity and help ensure successful experimental outcomes.
Primers and nucleotides are essential to every cDNA synthesis experiment. We offer predesigned and custom primers.
Value choice first strand cDNA synthesis kits for use with RT-PCR, RT-qPCR, and microarray applications.
Thermo Scientific offers a selection of reverse transcription reagents for robust performance at optimal value.
Discover several grades of nuclease-free water—diethylpyrocarbonate (DEPC)-treated water, nuclease-free water (not DEPC-treated), and RT-PCR grade water.
Resources
Reverse transcriptase webinars
Stay up-to-date on current industry trends in cDNA synthesis applications with on-demand webinars.
Reverse transcriptase literature
Access white papers and technical and application notes for insights on cutting edge reverse transcription technology.
Reverse transcriptase videos
Watch instructional videos on troubleshooting tips, how-to videos, and explainers related to cDNA synthesis.
Reverse transcriptase education
Review educational resources on topics including reverse transcription basics, cDNA synthesis, enzyme selection, troubleshooting tips, and cDNA applications in molecular biology.
Support
PCR and cDNA Synthesis Support Center
Find support for every step of your cDNA synthesis experiments.
Tm calculator
Handy tool to calculate Tm (melting temperature) of primers and annealing temperatures.
Contact us
Get in touch with our technical application scientists for questions regarding reverse transcription enzymes, master mixes, and cDNA synthesis kits.
Mobile and desktop applications
Access content and helpful tools through SmartIST and Cloning Bench mobile apps.
OEM and custom solutions
Source reverse transcriptase and other molecular biology enzymes, PCR reagents, and plastics customized to your needs.
For Research Use Only. Not for use in diagnostic procedures.