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Maxam-Gilbert Sequencing

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...

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Related Experiment Video

Updated: Jun 17, 2026

Assessment of DNase Activity by Ratiometric Fluorescence Resonance Energy Transfer

Assessment of DNase Activity by Ratiometric Fluorescence Resonance Energy Transfer

Published on: July 25, 2025

DNase I footprinting.

Antonia S Cardew¹, Keith R Fox

¹School of Biological Sciences, University of Southampton, Southampton, UK.

Methods in Molecular Biology (Clifton, N.J.)

|December 10, 2009

Summary

This summary is machine-generated.

DNA footprinting identifies DNA-binding compound sequence selectivity by observing how ligands shield DNA from cleavage. This method, using DNase I, helps determine compound binding sites and interaction strength.

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Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

DNA-binding compounds are crucial in various biological processes and therapeutic applications.
Understanding sequence selectivity is key to designing effective DNA-targeting agents.
Existing methods for assessing DNA-ligand interactions require refinement for accuracy and efficiency.

Purpose of the Study:

To detail the methodology of DNA footprinting for assessing DNA-binding compound sequence selectivity.
To provide a comprehensive guide on preparing DNA substrates and conducting footprinting experiments.
To explain how footprinting data can quantify the dissociation constant of DNA-ligand interactions.

Main Methods:

Preparation and labeling of DNA footprinting substrates (50-200 base pairs).
Performing DNA cleavage experiments using DNase I nuclease.
Analyzing cleavage patterns to identify DNA regions protected by ligands.

Main Results:

Demonstration of DNA footprinting as a reliable method for determining sequence selectivity.
Successful identification of ligand binding sites on DNA.
Quantification of DNA-ligand interaction strength through dissociation constant estimation.

Conclusions:

DNA footprinting is an effective technique for characterizing DNA-binding compounds.
The method provides insights into sequence-specific interactions critical for drug development.
Detailed protocols enable researchers to accurately estimate binding affinities.