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Products >  PCR_Products >  RT-PCR-Two_Step >  MMLV_Reverse_Transcriptase >  PrimeScript_High_Fidelity_RT-PCR_Kit

RT-PCR for Gene Expression: PrimeScript High Fidelity RT-PCR Kit

PrimeScript Reverse Transcriptase, included in the PrimeScript High Fidelity RT-PCR Kit, efficiently synthesizes cDNA thanks to its exceptionally strong strand displacement activity. This Reverse Transcriptase is robust, versatile and well-suited for applications requiring full-length cDNA, such as RT-PCR gene expression studies. PrimeScript High Fidelity Reverse Transcriptase can be used for reverse transcription of virtually any RNA template, including templates that are GC-rich and/or have high levels of secondary structure. PrimeScript RTase is a modified, recombinant RNase H Minus MMLV (Moloney Murine Leukemia Virus). Because of the excellent extension capability of this enzyme, cDNA synthesis can be performed at 42°C, decreasing the risk of RNA degradation that often occurs during conventional reactions performed at higher temperatures.

Fast and Accurate Quantification

During RT-PCR gene expression studies, accurate reverse transcription and PCR amplification are essential. The PrimeScript High Fidelity RT-PCR Kit includes all reagents needed for high fidelity amplification of cDNA from total RNA or mRNA and is designed for two-step endpoint RT-PCR. This kit includes PrimeScript High Fidelity Reverse Transcriptase, a highly accurate MMLV reverse transcriptase, and PrimeSTAR Max DNA Polymerase, which has the highest fidelity of any commercially available PCR enzyme. PrimeSTAR Max DNA Polymerase is formulated as a premix and offers high amplification efficiency as well as compatibility with a wide range of template concentrations. As a result, this kit is well suited for any cDNA studies requiring high fidelity, such as RT-PCR gene expression studies (including gene splicing studies where alternative splicing products must be distinguished, or when discerning between closely related gene family members), cDNA library construction, and transcriptome analysis.

First introduced in Japan, PrimeScript is now available to scientists worldwide. Over 3,700 peer-reviewed articles have been published in which PrimeScript High Fidelity Reverse Transcriptase is used for a variety of applications such as RT-PCR gene expression studies, transcriptome analyses, miRNA expression and regulation, molecular evolution, virology and microbiology.

  At-A-Glance   Documents   Images & Data   Resources

Features

  • Exceptionally low error rates during reverse transcription and PCR amplification
  • Excellent extension capacity even with highly structured template RNAs containing higher-order structure
  • Broadly tolerates a wide range of input RNA template concentrations
  • Applications

    • RT-PCR gene expression stydies
    • Full-length high fidelity cDNA synthesis and analysis by endpoint PCR (gel electrophoresis)
    • Two step RT-PCR
    • cDNA library construction

    Components

    R022A (50 reactions)*1

    PrimeScript High Fidelity Reverse Transcriptase (for two step RT-PCR)25 μL
    5 × PrimeScript Buffer200 μL
    RNase Inhibitor (40 U / μL)25 μL
    5 × PrimeScript Buffer200 μL
    dNTP Mixture (10 mM each)50 μL
    Oligo dT Primer (2.5 μM)50 μL
    Random 6 mers (20 μM)50 μL
    PrimeSTAR Max Premix (2×)625 μL × 2
    Control F-1 Primer*2(20 μM)10 μL
    R-Control Primer 1*3 (20 μM)10 μL
    Control RNA positive (2 × 105 copies / μL)20 μL
    RNase Free dH2O1 mL

    *1: For 50 reactions of 2-step RT-PCRs (20 μL RT reaction volume → 50 μL PCR volume)

    *2: Upstream sense primer for the Positive Control RNA

    *3: Downstream antisense primer for the Positive Control RNA

    R022B (200 reactions)

    This kit is comprised of 4 x R022A

    Storage

    -20°C

    Choosing a PrimeScript Kit for Endpoint RT-PCR

    If you want to....

    Use this....

    Prepare full-length cDNA using your own primers

    PrimeScript Reverse Transcriptase (Cat. # 2680A and 2680B)

    Prepare full-length cDNA using a kit that includes primers

    PrimeScript First Strand cDNA Synthesis Kit (Cat. # 6110A and 6110B)

    Perform 2-step RT-PCR with a kit that includes RTase, primers, and PCR enzyme

    PrimeScript RT-PCR Kit (Cat. # RR014A and RR014B)

    Perform cDNA cloning studies with extremely high fidelity

    PrimeScript High Fidelity RT-PCR Kit (Cat. # R022A and R022B)

    Perform 1-step PCR

    PrimeScript  One-Step RT-PCR Kit, Ver. 2 (Cat. # RR055A and RR055B)

    References

    1. Cui, X. et al. (2008) Acta Biochim. Biophys. Sin. 40:349-355.

    2. Matsumoto, T. et al. (2009) Plant Cell Physiol. 50:216-229.

    3. Qiu, L. et al. (2008) J. Biol. Chem. 283:17175-17183.

    4. Si, D. et al. (2012) Appl. Envir. Microbiol. 78:7923-7930.

    5. Tanaka, K. et al. (2012) Appl. Envir. Microbiol. 78:8161-8163.

    6. Wi, S. J. et al. (2012) Plant Physiol. 159:251-265.

    7. Wu, F. et al. (2011) Clin. Cancer Res. 17:2702-2711. 




     
     
    Products
    Cat. # Product Contents Size Price License Units Select
    R022A PrimeScript™ High Fidelity RT-PCR Kit 50 Rxns $281.00 License Statements
    R022B PrimeScript™ High Fidelity RT-PCR Kit 200 Rxns $796.00 License Statements
     

     

     
     

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