OneTaq® RT-PCR Kit

Easy and efficient two-step RT-PCR

  • Optimized reaction mixes for cDNA synthesis and PCR amplification
  • Hot start formulation eliminates nonspecific amplification and does not require a separate activation step
  • OneTaq® DNA Polymerase delivers robust amplification across a wide range of templates

Ordering Information

$name
  • 30 reactions
    $140.00
  • Product Information

    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 Synthesis Figure 1. First Strand DNA Synthesis

    First strand cDNA systhesis 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 2-Log DNA Ladder (NEB #N3200 ).
    Figure 2. First Strand DNA Synthesis Figure 2. First Strand DNA Synthesis


    Kit Components

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    Nuclease-free Water -20
    M-MuLV Enzyme Mix 10 X
    M-MuLV Reaction Mix 2 X
    OneTaq® Hot Start 2X Master Mix with Standard Buffer -20 2 X
    Oligo d(T)23 VN -20 50 μM
    Random Primer Mix -20 60 μM
    Product Categories:
    RT-PCR Products,
    Reverse Transcriptases & RT-PCR Products,
    cDNA Synthesis & Reverse Transcriptases
    Applications:
    RT-PCR,
    RT-PCR & cDNA Synthesis,
    cDNA Synthesis & RT-PCR
    ,
    Multiplex PCR
    ,
    Specialty PCR
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