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 Experiment Videos

Methylation-sensitive polymerase chain reaction.

Hannah R Moore1, Richard R Meehan, Lorraine E Young

  • 1Division of Gene Function and Development, Roslin Institute, United Kingdom.

Methods in Molecular Biology (Clifton, N.J.)
|June 10, 2006
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Correction: The potent human CAR activator CITCO is a non-genotoxic hepatic tumour-promoting agent in humanised constitutive androstane receptor mice but not in wild-type animals.

Archives of toxicology·2026
Same author

Letter to the editor regarding the review article: Yamada T, Cohen SM, Lake BG. The modes of action for rodent liver tumor formation by activators of the constitutive androstane receptor (CAR) and the peroxisome proliferator-activated receptor alpha (PPARα) are not relevant to human cancer risk: an updated critical evaluation. Crit Rev Toxicol. 2025;55(5):549-586. doi: 10.1080/10408444.2025.2513332.

Critical reviews in toxicology·2026
Same author

Loss of colonic fidelity enables multilineage plasticity and metastasis.

Nature·2025
Same author

The potent human CAR activator CITCO is a non-genotoxic hepatic tumour-promoting agent in humanised constitutive androstane receptor mice but not in wild-type animals.

Archives of toxicology·2025
Same author

Rett syndrome: interferon-γ to the rescue?

EMBO molecular medicine·2024
Same author

Neutrophils fuel effective immune responses through gluconeogenesis and glycogenesis.

Cell metabolism·2021

A new methylation-sensitive PCR method accurately measures DNA methylation in individual ovine embryos. This technique enables comparisons across various embryo production methods, offering a faster, cheaper alternative to bisulfite sequencing.

Area of Science:

  • Reproductive Biology
  • Epigenetics
  • Molecular Biology

Background:

  • Epigenetic modifications, such as DNA methylation, play crucial roles in early embryonic development.
  • Understanding methylation patterns in pre-implantation embryos is vital for assessing developmental competence and reproductive technologies.
  • Existing methods for DNA methylation analysis can be time-consuming, costly, or lack single-embryo resolution.

Purpose of the Study:

  • To develop and validate a robust and reproducible methylation-sensitive polymerase chain reaction (MS-PCR) method.
  • To quantify the percentage of DNA methylation in repeat sequences of individual pre-implantation ovine embryos.
  • To enable comparative analysis of methylation patterns in embryos produced via different biotechnologies.

Main Methods:

Related Experiment Videos

  • DNA extraction from single ovine embryos.
  • Digestion of DNA with a methylation-sensitive restriction enzyme.
  • Methylation-sensitive polymerase chain reaction (MS-PCR) amplification and quantification using image analysis of agarose gel electrophoresis.
  • Comparison with an undigested control representing 100% methylation.
  • Validation using a standard curve for individual embryo analysis.

Main Results:

  • The MS-PCR method reliably detects and quantifies DNA methylation percentage in individual pre-implantation ovine embryos.
  • The method accounts for inter-embryo heterogeneity within treatment groups.
  • Results are validated against a standard curve for accuracy.
  • The MS-PCR approach demonstrates significant advantages in speed, cost, and throughput compared to bisulfite sequencing.

Conclusions:

  • The described MS-PCR method is a robust, reproducible, and efficient tool for analyzing DNA methylation in single embryos.
  • This technique facilitates the comparison of epigenetic profiles across various embryo production methods, including nuclear transfer.
  • The MS-PCR method offers a practical and high-throughput alternative for epigenetic studies in early-stage embryos.