OneTaq® RT-PCR Kit combines two powerful mixes, M-MuLV Enzyme Mix and OneTaq Hot Start 2X Master Mix with Standard Buffer for 2-step RT-PCR applications. The two mixes require minimal handling during reaction setup and yet offer consistent and robust RT-PCR reactions.The first strand cDNA synthesis is achieved by using two optimized mixes, M-MuLV Enzyme Mix and M-MuLV Reaction Mix. M-MuLV Enzyme Mix combines M-MuLV Reverse Transcriptase and murine RNase Inhibitor while M-MuLV Reaction Mix contains dNTPs and an optimized buffer. The kit also contains two optimized primers for reverse transcription and nuclease-free water. An anchored oligo-dT primer [d(T)23VN] forces the primer to anneal to the beginning of the polyA tail. The optimized Random Primer Mix provides random and consistent priming sites covering the entire RNA templates including both mRNAs and non-polyadenylated RNAs.The amplification step features a OneTaq Hot Start DNA Polymerase in a master mix format. OneTaq Hot Start DNA Polymerase offers higher fidelity than Taq and better amplification. RT-PCR product up to 6 kb can be generated (Figure 1).
Figure 1. First Strand DNA SynthesisFirst strand cDNA synthesis was carried out in the presence of 1X M-MuLV Enzyme Mix at 42°C using 0.5 μg of human spleen total RNA in the presence of dT23VN (lanes 1, 4 and 7) or Random Hexamer Mix (lanes 2, 5 and 8). No-RT controls were lanes 3, 6 and 9. OneTaq Hot Start 1X Master Mix was used to amplify a 1.5 kb fragment of beta-actin gene, a 0.6 kb fragment of GAPDH gene, and a 5.5 kb fragment from p532 gene in 35 cycles. The marker lane (M) contains 1 kb Plus DNA Ladder (NEB #N3200 ).Figure 2. First Strand DNA Synthesis
This product is related to the following categories:
RT-PCR Products,
Reverse Transcriptases & RT-PCR Products,
cDNA Synthesis & Reverse Transcriptases Products
This product can be used in the following applications: