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Related Concept Videos

Labeling DNA Probes03:31

Labeling DNA Probes

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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
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Related Experiment Video

Updated: Aug 24, 2025

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution
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Post-bisulfite Adaptor Tagging with a Highly Efficient Single-Stranded DNA Ligation Technique.

Fumihito Miura1, Takashi Ito2

  • 1Department of Biochemistry, Kyushu University Graduate School of Medical Sciences, Fukuoka, Japan. fumihito@med.kyushu-u.ac.jp.

Methods in Molecular Biology (Clifton, N.J.)
|October 20, 2022
PubMed
Summary

The tPBAT protocol improves whole-genome bisulfite sequencing library preparation. This enhanced method increases read length and mapping rates, overcoming limitations of the original post-bisulfite adaptor tagging procedure.

Keywords:
DNA methylationMethylomePost-bisulfite adaptor tagging (PBAT)Single-stranded DNA ligationTACS ligationWhole-genome bisulfite sequencing (WGBS)

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Area of Science:

  • Molecular Biology
  • Genomics
  • Epigenetics

Background:

  • Post-bisulfite adaptor tagging (PBAT) enables efficient library preparation for whole-genome bisulfite sequencing (WGBS).
  • The original PBAT protocol excelled with limited sample amounts but had limitations.
  • Two random priming steps in the original PBAT hindered improvements in read length and mapping rates.

Purpose of the Study:

  • To describe an improved version of the PBAT protocol, termed tPBAT.
  • To address the limitations of the original PBAT protocol regarding read length and mapping efficiency.

Main Methods:

  • The chapter details the tPBAT protocol, an advancement over the original PBAT.
  • Key modifications focus on optimizing the library preparation process for WGBS.

Main Results:

  • The tPBAT protocol is designed to enhance sequencing library preparation efficiency.
  • Improvements are expected in read length and mapping rates compared to the original PBAT.

Conclusions:

  • The tPBAT protocol offers a significant advancement in WGBS library preparation.
  • This improved method facilitates more accurate and comprehensive epigenetic analysis.