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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
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Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRAC
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Published on: May 9, 2020

Real-Time RT-PCR: cDNA Synthesis.

Wolfgang Kusser, Sandrine Javorschi, Martin A Gleeson

    CSH Protocols
    |April 10, 2012
    PubMed
    Summary
    This summary is machine-generated.

    This study details a protocol for creating complementary DNA (cDNA) from total RNA using the Superscript II First-Strand Synthesis System. This method is crucial for downstream molecular biology applications involving gene expression analysis.

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    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Genetics

    Background:

    • Complementary DNA (cDNA) synthesis is a fundamental technique in molecular biology.
    • Accurate cDNA generation is essential for studying gene expression and RNA sequencing.
    • Various methods exist for first-strand cDNA synthesis, each with specific advantages.

    Purpose of the Study:

    • To describe a reliable protocol for first-strand cDNA synthesis.
    • To outline the application of the Superscript II First-Strand Synthesis System.
    • To provide a foundation for subsequent molecular analyses.

    Main Methods:

    • Utilized the Superscript II First-Strand Synthesis System.
    • Employed total RNA as the starting material.
    • Followed a standardized protocol for cDNA generation.

    Main Results:

    • Successfully generated cDNA from total RNA.
    • The protocol yielded high-quality cDNA suitable for downstream applications.
    • Demonstrated the efficacy of the Superscript II system for this purpose.

    Conclusions:

    • The Superscript II First-Strand Synthesis System provides an effective method for cDNA synthesis from total RNA.
    • This protocol is valuable for researchers in molecular biology and genomics.
    • The generated cDNA can be used for various applications, including gene expression studies.