Related Experiment Video
Updated: Jul 10, 2026
08:23
Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
Published on: September 25, 2018
[Methodological evaluation on PCR-DGGE technique in detecting DNA mutation and single nucleotide polymorphism]
Xiao-jun Lu1, Yong-qian Jia, Hong Fan
1Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Summary
Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) effectively detects DNA mutations and single nucleotide polymorphisms (SNPs) with high accuracy. This method shows excellent sensitivity and specificity for genetic variation analysis.
Area of Science:
- Molecular Biology
- Genetics
Context:
- Acute leukemia patients' mitochondrial DNA D-loop mutations were analyzed.
- DNA sequencing and PCR-DGGE were compared for mutation detection efficiency.
Purpose:
- To evaluate the efficiency of PCR-DGGE in identifying DNA base mutations and SNPs.
- To assess the sensitivity of PCR-DGGE for detecting DNA polymorphism.
Summary:
- PCR-DGGE demonstrated 100% specificity and 97.1% sensitivity for base mutation detection.
- The technique could detect polymorphism down to a 0.5% mixture of mutated DNA.
- Results were compared against DNA sequencing for validation.
Impact:
- PCR-DGGE is a highly sensitive and specific method for detecting DNA mutations and SNPs.
- This technique offers a valuable tool for genetic variation analysis in clinical and research settings.
Related Concept Videos
01:32PCR - Polymerase Chain Reaction
Overview
01:32PCR
Overview
01:57Sanger Sequencing
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
01:24DNA Isolation
DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...