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Related Concept Videos

Proofreading01:31

Proofreading

Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore,  it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase Enzyme
Proofreading01:43

Proofreading

Synthesis of new DNA molecules starts when DNA polymerase links nucleotides together in a sequence that is complementary to the template DNA strand. DNA polymerase has a higher affinity for the correct base to ensure fidelity in DNA replication. The DNA polymerase furthermore proofreads during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.Errors during Replication Are Corrected by the DNA Polymerase EnzymeGenomic DNA is synthesized in...
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
The Replisome03:01

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
The Replisome03:01

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
Homologous Recombination02:31

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

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Related Experiment Video

Updated: Jun 28, 2026

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
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DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

Published on: October 6, 2017

RNA-dependent DNA polymerases.

George Tzertzinis, Stanley Tabor, Nicole M Nichols

    Current Protocols in Molecular Biology
    |October 31, 2008
    PubMed
    Summary
    This summary is machine-generated.

    This study details using reverse transcriptases (RTs) for complementary DNA (cDNA) synthesis from RNA templates. It covers reaction conditions for Moloney murine leukemia virus (MMLV) and avian myeloblastosis virus (AMV) RTs, enabling applications like PCR and DNA sequencing.

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    Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes

    Published on: April 4, 2025

    Area of Science:

    • Molecular Biology
    • Enzymology

    Background:

    • Reverse transcriptases (RTs) are key enzymes that synthesize DNA from RNA templates.
    • They are primarily utilized in molecular biology for complementary DNA (cDNA) synthesis, a crucial step for various downstream applications.
    • Oligodeoxynucleotides serve as primers for initiating DNA synthesis on RNA templates.

    Purpose of the Study:

    • To describe optimal reaction conditions for Moloney murine leukemia virus (MMLV) and avian myeloblastosis virus (AMV) reverse transcriptases.
    • To outline diverse applications of RTs, including cDNA synthesis, 3' end labeling, and DNA sequencing.
    • To provide a comprehensive guide for utilizing RTs in molecular biology workflows.

    Main Methods:

    • Detailed protocols for reverse transcription reactions using MMLV and AMV RTs.
    • Methods for synthesizing cDNA from RNA templates.
    • Techniques for 3' fill-in reactions and labeling DNA fragments.

    Main Results:

    • Established appropriate reaction conditions for MMLV and AMV RTs.
    • Demonstrated the utility of RTs in synthesizing cDNA for PCR and cloning.
    • Showcased applications in labeling DNA termini and DNA sequencing.

    Conclusions:

    • Reverse transcriptases are versatile enzymes with critical roles in molecular biology.
    • Optimized reaction conditions enhance the efficiency of cDNA synthesis and other enzymatic activities.
    • RTs facilitate diverse applications, from gene expression analysis to DNA fragment manipulation.