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DNA Isolation

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...

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Updated: Feb 23, 2026

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

Published on: June 6, 2020

Rapid Gene Isolation Using MutChromSeq.

Burkhard Steuernagel¹, Jan Vrána², Miroslava Karafiátová²

¹John Innes Centre, Norwich Research Park, Norwich, UK.

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

|September 1, 2017

Summary

This summary is machine-generated.

MutChromSeq isolates genes controlling plant gene expression in complex genomes. This method uses chromosome sorting and sequencing to identify mutations without genetic mapping.

Keywords:

Chromosome isolation FISHIS Flow cytometry and chromosome sorting Gene cloning Mutational genomics

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Area of Science:

Plant genomics
Molecular biology
Genetics

Background:

Plants with complex and polyploid genomes present challenges for gene discovery.
Identifying genes that control gene expression is crucial for understanding plant biology and breeding.

Purpose of the Study:

To introduce MutChromSeq, a novel approach for isolating genes and DNA sequences that control gene expression.
To enable efficient gene discovery in plants with complex and polyploid genomes.

Main Methods:

MutChromSeq employs lossless complexity reduction through flow cytometric chromosome sorting.
DNA from isolated chromosomes undergoes shotgun sequencing.
Comparative sequence analysis between wild-type and mutant chromosomes identifies causative mutations.

Main Results:

MutChromSeq successfully identifies causative mutations in single candidate genes or noncoding sequences.
The approach is effective for plants with complex and polyploid genomes.
Mutations can be identified without relying on recombination-based genetic mapping.

Conclusions:

MutChromSeq provides a powerful tool for gene isolation and mutation identification in challenging plant genomes.
This method expands the scope of targeted DNA sequences beyond traditional genetic mapping limitations.