Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Histone Variants at the Centromere02:30

Histone Variants at the Centromere

5.1K
Histone variants are the histone proteins with structural and sequence variations. These variants may be regarded as “mutant” forms that replace their canonical histone counterparts in the nucleosomes. Specific post-translational modifications on the histone variants enable further chromatin complexity and regulate tissue-specific gene expression. The most common histone variants are from histone H2A, H2B, and linker histone H1 families. However, several variants of histone H3...
5.1K
PCR01:32

PCR

237.4K
Overview
237.4K
Real Time RT-PCR02:57

Real Time RT-PCR

65.1K
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...
65.1K
Transmission-based Precautions I: Contact, Enteric, and Droplets01:17

Transmission-based Precautions I: Contact, Enteric, and Droplets

4.6K
Transmission-based precautions are for patients known to be infected or suspected to be infected or colonized with organisms that pose a significant risk to others. Some transmission-based precautions include contact, enteric, and droplet.
Contact Precautions:
Contact precautions are the measures taken to prevent the transmission of infectious agents, especially epidemiologically important microorganisms such as MRSA or influenza, primarily transmitted through direct or indirect contact with an...
4.6K
Quantitative Analysis01:12

Quantitative Analysis

1.4K
Quantitative analysis is a technique for measuring the amount of specific constituents in a sample. When the sample's composition is unknown, qualitative analysis is performed first to identify its components, which ensures that the correct substances are measured during the quantitative phase.
In quantitative analysis, two key measurements are made: the sample quantity and a property proportional to the amount of the analyte (the substance being analyzed). This forms the basis of the...
1.4K
Social Foundations of Self IV: Self in Digital Communication01:30

Social Foundations of Self IV: Self in Digital Communication

202
Since the early 2000s, computer-mediated communication (CMC) has grown rapidly, playing a crucial role in self-development. A key distinction between CMC and real-life interactions is the lack of a physically present partner. This absence makes non-verbal cues such as facial expressions, body language, and paralinguistic signals unavailable in CMC platforms like email, instant messaging, or social media. The lack of these cues can create ambiguity and complicate how feedback is interpreted.The...
202

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Heterogeneity of neutrophils and inflammatory responses in patients with COVID-19 and healthy controls.

Frontiers in immunology·2022
Same author

Pharmacological inhibition of Carbonic Anhydrase IX and XII to enhance targeting of acute myeloid leukaemia cells under hypoxic conditions.

Journal of cellular and molecular medicine·2021
Same author

Type 1 interferon to prevent leukemia relapse after allogeneic transplantation.

Blood advances·2021
Same author

Therapeutic targeting of TP53-mutated acute myeloid leukemia by inhibiting HIF-1α with echinomycin.

Oncogene·2020
Same author

A Phase 2 Study of Pembrolizumab during Lymphodepletion after Autologous Hematopoietic Cell Transplantation for Multiple Myeloma.

Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation·2019
Same author

Maintenance sorafenib in FLT3-ITD AML following allogeneic HCT favorably impacts relapse and overall survival.

Bone marrow transplantation·2019

Related Experiment Video

Updated: Feb 3, 2026

Detection of a CDH1 Rare Transcript Variant in Fresh-frozen Gastric Cancer Tissues by Chip-based Digital PCR
09:16

Detection of a CDH1 Rare Transcript Variant in Fresh-frozen Gastric Cancer Tissues by Chip-based Digital PCR

Published on: February 5, 2018

6.6K

Rare Variant Quantitation Using Droplet Digital PCR.

Brian Parkin1,2

  • 1Division of Hematology and Oncology, Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA. bparkin@med.umich.edu.

Methods in Molecular Biology (Clifton, N.J.)
|October 24, 2018
PubMed
Summary
This summary is machine-generated.

Droplet digital PCR (ddPCR) precisely quantifies DNA copies using allele-specific probes. This method achieves highly sensitive detection of mutant genes in genomic DNA, down to 1 in 50,000 copies.

Keywords:
Allele-specific probesDigital PCRLocked nucleic acidMutation quantitationPolymerase chain reactionRare variant detection

More Related Videos

Quantification of Circular RNAs Using Digital Droplet PCR
08:39

Quantification of Circular RNAs Using Digital Droplet PCR

Published on: September 16, 2022

4.1K
Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in Murine Retina
09:50

Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in Murine Retina

Published on: December 25, 2021

5.2K

Related Experiment Videos

Last Updated: Feb 3, 2026

Detection of a CDH1 Rare Transcript Variant in Fresh-frozen Gastric Cancer Tissues by Chip-based Digital PCR
09:16

Detection of a CDH1 Rare Transcript Variant in Fresh-frozen Gastric Cancer Tissues by Chip-based Digital PCR

Published on: February 5, 2018

6.6K
Quantification of Circular RNAs Using Digital Droplet PCR
08:39

Quantification of Circular RNAs Using Digital Droplet PCR

Published on: September 16, 2022

4.1K
Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in Murine Retina
09:50

Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in Murine Retina

Published on: December 25, 2021

5.2K

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Precise DNA quantitation is crucial for genetic analysis.
  • Detecting low-frequency mutations requires highly sensitive methods.
  • Droplet digital PCR (ddPCR) offers advanced capabilities for nucleic acid quantification.

Purpose of the Study:

  • To describe a ddPCR-based protocol for detecting mutations in genomic DNA.
  • To establish a highly sensitive method for variant allele frequency (VAF) quantitation.
  • To demonstrate the utility of ddPCR for sensitive mutation detection.

Main Methods:

  • Utilized Droplet digital PCR (ddPCR) technology.
  • Employed allele-specific fluorescent oligonucleotide probes for discrimination.
  • Developed a protocol for enumerating mutant and wild-type alleles.
  • Applied the method to genomic DNA samples.

Main Results:

  • Achieved precise quantitation of individual DNA copies.
  • Enabled sensitive detection and quantification of variant allele frequencies.
  • Demonstrated a detection sensitivity of up to 1 in 50,000 DNA copies for mutations.
  • Successfully enumerated both mutant and wild-type alleles.

Conclusions:

  • ddPCR provides a robust platform for sensitive mutation detection in genomic DNA.
  • The described protocol enables accurate quantitation of low-frequency genetic variants.
  • This method significantly enhances the capability for identifying mutations with high precision.