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Related Experiment Video

Updated: Dec 11, 2025

DNA Methylation: Bisulphite Modification and Analysis
12:34

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

106.0K

Hairpin-Bisulfite PCR.

Reinhard Stöger1

  • 1School of Biosciences, University of Nottingham, Nottingham, UK. reinhard.stoger@nottingham.ac.uk.

Methods in Molecular Biology (Clifton, N.J.)
|August 22, 2020
PubMed
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This study introduces hairpin bisulfite sequencing, a method that links DNA strands to reveal cytosine methylation dynamics on both strands of individual DNA molecules, overcoming conventional sequencing limitations.

Area of Science:

  • Epigenetics
  • Molecular Biology
  • Genomics

Background:

  • Conventional bisulfite sequencing analyzes only one DNA strand, limiting epigenetic analysis.
  • Understanding DNA methylation dynamics on both strands is crucial for epigenetic memory research.

Purpose of the Study:

  • To present a novel hairpin bisulfite sequencing method for analyzing DNA methylation on both strands of individual DNA molecules.
  • To provide a detailed protocol for generating hairpin-linked DNA and performing efficient bisulfite conversion.

Main Methods:

  • Ligation of a hairpin oligonucleotide to genomic DNA.
  • Bisulfite conversion of DNA.
  • Polymerase chain reaction (PCR) amplification.
  • Sequencing of hairpin bisulfite PCR products.
Keywords:
Ammonium sulfite monohydrateBisulfite conversionBisulfite sequencingCpG dyadDNA methylationDouble-stranded DNAEpigeneticsHairpin bisulfite PCR

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Related Experiment Videos

Last Updated: Dec 11, 2025

DNA Methylation: Bisulphite Modification and Analysis
12:34

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

106.0K
Single Oocyte Bisulfite Mutagenesis
13:18

Single Oocyte Bisulfite Mutagenesis

Published on: June 27, 2012

14.3K
Methodology for Accurate Detection of Mitochondrial DNA Methylation
12:11

Methodology for Accurate Detection of Mitochondrial DNA Methylation

Published on: May 20, 2018

13.8K

Main Results:

  • The hairpin ligation physically links complementary DNA strands.
  • This method enables the analysis of cytosine methylation status on both strands simultaneously.
  • Sequences from hairpin bisulfite PCR products reveal epigenetic dynamics on individual DNA molecules.

Conclusions:

  • Hairpin bisulfite sequencing overcomes limitations of conventional methods by analyzing both DNA strands.
  • The described procedure offers a reliable approach for studying epigenetic memory systems.
  • The protocol is suitable for both conventional and hairpin bisulfite sequencing applications.