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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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Related Experiment Video

Updated: May 15, 2025

Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing

Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing

Published on: March 31, 2017

High Molecular Weight DNA Extraction from Wheat Rust Spores.

Meng Li¹, Gurcharn Singh Brar^1,2, Benjamin Schwessinger³

¹Faculty of Land and Food Systems, The University of British Columbia, Vancouver, BC, Canada.

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

|April 8, 2025

Summary

Researchers developed a new DNA extraction method to obtain high molecular weight (HMW) DNA from wheat rust spores. This breakthrough enables advanced long-read sequencing for better understanding of rust genetics and evolution.

Keywords:

CTAB DNA purification HMW DNA Rust Stripe rust Urediniospore

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Last Updated: May 15, 2025

Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing

Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing

Published on: March 31, 2017

Extraction of High Molecular Weight Genomic DNA from Soils and Sediments

Extraction of High Molecular Weight Genomic DNA from Soils and Sediments

Published on: November 10, 2009

Extraction of High Molecular Weight DNA from Microbial Mats

Extraction of High Molecular Weight DNA from Microbial Mats

Published on: July 7, 2011

Area of Science:

Plant Pathology
Genomics
Molecular Biology

Background:

Wheat rust diseases pose a significant threat to global food security due to rapid spread and evolution.
Dikaryotic urediniospores of rust fungi present challenges for genomic analysis using short-read sequencing.
Understanding rust genome architecture is crucial for developing durable resistance in wheat cultivars.

Purpose of the Study:

To develop an improved DNA extraction protocol for wheat rust spores.
To obtain high molecular weight (HMW) DNA suitable for long-read sequencing.
To facilitate deeper insights into rust genetic variation and evolution.

Main Methods:

Optimization of DNA extraction from wheat rust spores/urediniospores.
Isolation of pure high molecular weight (HMW) DNA.
Assessment of DNA quality and quantity for downstream sequencing applications.

Main Results:

A substantial yield of pure HMW DNA was achieved from rust spores.
The isolated DNA averaged over 50 kilobase pairs (kbp) in molecular weight.
The protocol provides DNA suitable for long-read sequencing technologies (e.g., PacBio, Nanopore).

Conclusions:

The improved protocol overcomes challenges in HMW DNA extraction from rust spores.
This advancement enables high-resolution genome sequencing of wheat rust fungi.
Facilitates detailed exploration of rust genetic diversity, evolution, and virulence mechanisms.