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.
Finally, reverse transcription of low expression level RNA is also possible due to a 20 µL reverse transcription reaction in which a maximum RNA sample volume of 9.5 µL can be added. Takara Ex Taq HS (a high fidelity, hot start PCR polymerase), is used as the enzyme for cDNA amplification. This enzyme prevents non-specific DNA amplification due to mispriming and primer dimerization that may occur when preparing reaction solutions prior to cycling. Amplification of cDNA up to 5 kb in length is possible.
