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.

Cat. #	Product	Size	Price
RR019A	RNA PCR Kit (AMV) Version 3.0	100 Rxns	USD $540.00
The RNA PCR Kit (AMV) Version 3.0 is designed for performing single tube reverse transcription of RNA to cDNA using AMV (Avian Myeloblastosis Virus) Reverse Transcriptase, with subsequent cDNA amplification using Takara Ex Taq HS. Taking advantage of AMV RTase XL, which possesses higher thermostability and a broader range of reaction temperatures (42–60°C) than MMLV RTase, this kit allows transcription at higher temperatures, which eliminates potential RNA secondary structure that can give rise to polymerase pauses. The supplied Oligo dT-Adaptor Primer is designed for highly efficient cDNA synthesis from the poly(A)⁺ RNA 3'-terminus, which allows amplification of unknown 3'-termini using 3'-RACE. The components for the 3'-RACE method are supplied in the kit. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components You May Also Like Image Data Back Principle of RNA PCR Kit, Version 3.0 Principle of RNA PCR Kit, Version 3.0. Back RR019A: RNA PCR Kit (AMV) Version 3.0
RR019B	RNA PCR Kit (AMV) Version 3.0	200 Rxns	USD $1050.00
The RNA PCR Kit (AMV) Version 3.0 is designed for performing single tube reverse transcription of RNA to cDNA using AMV (Avian Myeloblastosis Virus) Reverse Transcriptase, with subsequent cDNA amplification using Takara Ex Taq HS. Taking advantage of AMV RTase XL, which possesses higher thermostability and a broader range of reaction temperatures (42-60°C) than MMLV RTase, this kit allows transcription at higher temperatures, which eliminates potential RNA secondary structure that can give rise to polymerase pauses. The supplied Oligo dT-Adaptor Primer is designed for highly efficient cDNA synthesis from the poly(A)⁺ RNA 3'-terminus, which allows amplification of unknown 3'-termini using 3'-RACE. The components for the 3'-RACE method are supplied in the kit. Cat. # RR019B contains 2 of Cat. # RR019A. Please refer to Cat. # RR019A for complete product documentation and resources. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents You May Also Like

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.

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