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Related Concept Videos

In vitro Mutagenesis01:16

In vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Related Experiment Video

Updated: May 20, 2026

Single Oocyte Bisulfite Mutagenesis
13:18

Single Oocyte Bisulfite Mutagenesis

Published on: June 27, 2012

Single oocyte bisulfite mutagenesis.

Michelle M Denomme1, Liyue Zhang, Mellissa R W Mann

  • 1Department of Obstretrics & Gynaecology, Schulich School of Medicine and Dentistry, University of Western Ontario, Ontario, Canada.

Journal of Visualized Experiments : Jove
|July 12, 2012
PubMed
Summary
This summary is machine-generated.

This study presents a new method for analyzing DNA methylation in single oocytes and early embryos. The improved bisulfite mutagenesis protocol protects minute DNA amounts, enabling precise epigenetic analysis at the single-cell level.

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Last Updated: May 20, 2026

Single Oocyte Bisulfite Mutagenesis
13:18

Single Oocyte Bisulfite Mutagenesis

Published on: June 27, 2012

Meiotic Spindle Assessment in Mouse Oocytes by siRNA-mediated Silencing
09:16

Meiotic Spindle Assessment in Mouse Oocytes by siRNA-mediated Silencing

Published on: October 11, 2015

Using Mouse Oocytes to Assess Human Gene Function During Meiosis I
11:13

Using Mouse Oocytes to Assess Human Gene Function During Meiosis I

Published on: April 10, 2018

Area of Science:

  • Epigenetics and molecular biology
  • Developmental biology
  • Genomics and bioinformatics

Background:

  • Epigenetics regulates gene transcription via heritable chromatin modifications.
  • DNA methylation, a key epigenetic mark, is crucial for development and gene silencing.
  • Existing bisulfite mutagenesis methods struggle with minute DNA amounts in single oocytes.

Purpose of the Study:

  • To develop a sensitive and reproducible method for DNA methylation analysis in individual oocytes and cleavage-stage embryos.
  • To overcome challenges of DNA degradation and loss in single-cell epigenetic studies.
  • To enable accurate nucleotide-level DNA methylation profiling from single cells.

Main Methods:

  • Developed a novel protocol involving direct embedding of oocytes in agarose and lysis solution beads after zona pellucida removal.
  • Utilized bisulfite mutagenesis to convert unmethylated cytosines to uracils, detected as thymines after PCR and sequencing.
  • Protected minute DNA quantities from degradation and loss throughout the protocol.

Main Results:

  • Successfully analyzed DNA methylation at the single-cell level in oocytes and cleavage-stage embryos.
  • Eliminated PCR bias inherent in pooled samples by analyzing individual cells.
  • Enabled detection of cumulus cell contamination by excluding samples with multiple methylation patterns.

Conclusions:

  • The new protocol offers a robust and reproducible method for single-cell DNA methylation analysis.
  • This advancement is critical for studying epigenetic regulation in early development.
  • The method enhances the sensitivity and accuracy of epigenetic studies on oocytes and early embryos.