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Sanger Sequencing

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DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...

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Reusable Single Cell for Iterative Epigenomic Analyses

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Published on: February 11, 2022

Single DNA molecule patterning for high-throughput epigenetic mapping.

Aline Cerf¹, Benjamin R Cipriany, Jaime J Benítez

¹School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, United States.

Analytical Chemistry

|October 11, 2011

Summary

This summary is machine-generated.

This study introduces a new method to map 5-methyl cytosine distribution on single DNA molecules using soft-lithography and fluorescent peptide binding. This technique enables high-resolution imaging of DNA methylation states.

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A Semiautomated ChIP-Seq Procedure for Large-scale Epigenetic Studies

A Semiautomated ChIP-Seq Procedure for Large-scale Epigenetic Studies

Published on: August 13, 2020

Area of Science:

Epigenetics and Molecular Biology
Biotechnology and Nanotechnology

Background:

DNA methylation is crucial for gene regulation.
Accurate profiling of 5-methyl cytosine on single DNA molecules is challenging.
Existing methods lack resolution or scalability.

Purpose of the Study:

To develop a novel method for high-resolution mapping of 5-methyl cytosine distribution on individual DNA molecules.
To enable precise analysis of DNA methylation patterns at the single-molecule level.

Main Methods:

Combines soft-lithography and molecular elongation to create ordered arrays of immobilized DNA molecules.
Utilizes fluorescently labeled methyl-CpG binding domain peptides for detection.
Employs high-resolution imaging of peptide distribution on elongated DNA.

Main Results:

Successfully generated ordered arrays with over 250,000 DNA molecules.
Demonstrated the ability to detect and map 5-methyl cytosine distribution on single DNA molecules.
Confirmed the technique's capability to image DNA molecules with varying methylation states.

Conclusions:

The developed method offers a powerful tool for single-molecule DNA methylation profiling.
This technique provides high spatial resolution and signal averaging for accurate analysis.
It opens new avenues for studying epigenetic modifications in various biological contexts.