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Related Concept Videos

Mismatch Repair01:20

Mismatch Repair

Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
Mismatch Repair01:36

Mismatch Repair

Overview
Spontaneous and Induced Mutations01:30

Spontaneous and Induced Mutations

Spontaneous mutations arise infrequently during DNA replication due to errors in the process. A key factor behind these errors is tautomeric shifts in nitrogenous bases, where bases transition from keto to enol forms or amino to imino forms. This shift can alter base-pairing rules, leading to mutations. Additionally, reactive oxygen species (ROS) arising from aerobic metabolism can damage DNA, resulting in depurination (loss of a purine base) or depyrimidination (loss of a pyrimidine base).
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...
Mutations in Microorganisms01:18

Mutations in Microorganisms

Mutations are heritable changes in an organism’s genome involving alterations in the base sequence of DNA or RNA. These changes can influence cellular processes and phenotypic traits, potentially transforming the unaltered wild type into a mutant form. Such changes, termed forward mutations, are pivotal in shaping the genetic diversity of organisms.RNA viruses exhibit the highest mutation rates due to the absence of robust proofreading mechanisms during genome replication. In contrast,...
Genome Copying Errors02:46

Genome Copying Errors

DNA replication is a well-evolved process that copies millions of base pairs with high fidelity during each cell division. Occasionally a wrong base or a long stretch of wrong bases may get added to the daughter strands. If the errors are left unchecked, cells might accumulate several mutations that might endanger their  survival. Therefore, the copying errors are checked and repaired at three levels.

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Related Experiment Video

Updated: Jun 28, 2026

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations
10:41

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Published on: March 29, 2017

Single molecule PCR in mtDNA mutational analysis: Genuine mutations vs. damage bypass-derived artifacts.

Y Kraytsberg1, A Nicholas, P Caro

  • 1Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, MA 02215, USA.

Methods (San Diego, Calif.)
|October 29, 2008
PubMed
Summary
This summary is machine-generated.

Somatic mitochondrial DNA (mtDNA) mutation quantification is unreliable due to PCR artifacts. Single molecule PCR (smPCR) offers a novel solution to accurately measure mtDNA mutations, addressing challenges with polymerase bypass errors.

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Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
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Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

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Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
07:24

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

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Related Experiment Videos

Last Updated: Jun 28, 2026

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations
10:41

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Published on: March 29, 2017

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
08:23

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
07:24

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

Area of Science:

  • Mitochondrial genetics
  • Molecular biology
  • Genomics

Background:

  • Somatic mitochondrial DNA (mtDNA) mutation measurement methods yield highly variable results, spanning orders of magnitude.
  • The discrepancies are suspected to stem from Polymerase Chain Reaction (PCR) artifacts, including polymerase errors and bypass of damaged nucleotides.
  • Current methods lack adequate controls to account for bypass-related artificial mutations.

Purpose of the Study:

  • To address the crisis in somatic mtDNA mutation measurement.
  • To propose single molecule PCR (smPCR) as a method to account for artificial mutations.
  • To discuss the methodology of using smPCR for accurate mtDNA mutation quantification.

Main Methods:

  • Utilizing single molecule PCR (smPCR) to isolate and amplify individual mtDNA molecules.
  • Implementing controls within smPCR to differentiate true mutations from PCR-induced artifacts.
  • Analyzing the fidelity of DNA polymerases in the context of damaged nucleotide bypass.

Main Results:

  • smPCR is uniquely capable of identifying and quantifying artificial mutations arising from polymerase bypass of damaged nucleotides.
  • This technique provides a robust framework for controlling for specific PCR-related artifacts.
  • The methodology allows for more accurate measurement of endogenous somatic mtDNA mutations.

Conclusions:

  • Single molecule PCR (smPCR) is essential for resolving discrepancies in somatic mtDNA mutation quantification.
  • This method provides a critical tool for understanding the true rate of mtDNA mutagenesis.
  • Accurate mtDNA mutation measurement using smPCR will advance research in aging and disease.