Two-step RT-PCR
The RNA PCR Kit (AMV) Version 3.0 combines AMV Reverse Transcriptase with Takara Ex Taq HS—a high-fidelity, hot-start DNA polymerase—enabling users to synthesize and amplify cDNA in a single tube, using a two-step protocol. The RNA PCR kit uses AMV RTase XL, which possesses higher thermostability and a broader range of reaction temperatures (42–60°C) than MMLV RT. This allows transcription to occur at higher temperatures and helps eliminate potential RNA secondary structures that can impede downstream cDNA amplification. The supplied Oligo (dT) Adaptor primer is designed for highly efficient cDNA synthesis from the poly A+ RNA 3'-terminus, allowing later amplification of unknown 3'-termini using 3'-RACE. Components for 3'-RACE are supplied in this two-step RT-PCR kit.
More Information
Applications
- Two-step RT-PCR
- cDNA synthesis
Notes
Primers and Positive control RNA are the same as those supplied in the RNA LA PCR Kit.
Reverse transcription of RNA expressed at low levels (rare RNAs) may also be performed by creating a 20 µl reaction mixture with an RNA sample volume of up to 9.5 µl. Takara Ex Taq HS is the enzyme of choice for cDNA amplification because it prevents non-specific room temperature DNA amplification due to mispriming and primer dimerization. Amplification of cDNAs up to 5 kb in length is possible using this RNA PCR kit.
Product citations
Frohman, M. A., Dush, M. K. & Martin, G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc. Natl. Acad. Sci. U. S. A. 85, 8998–9002 (1988).
Kawasaki, E. S., and Wang, A. M. PCR Technology: Principles and Applications for DNA Amplification. (Palgrave Macmillan UK, 1989).
Lynas, C., Cook, S. D., Laycock, K. A., Bradfield, J. W. & Maitland, N. J. Detection of latent virus mRNA in tissues using the polymerase chain reaction. J. Pathol. 157, 285–9 (1989).
Additional product information
Please see the product’s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
Takara Bio USA, Inc.
United States/Canada: +1.800.662.2566 • Asia Pacific: +1.650.919.7300 • Europe: +33.(0)1.3904.6880 • Japan: +81.(0)77.565.6999
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. © 2025 Takara Bio Inc. All Rights Reserved. All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at takarabio.com.
