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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: Jul 23, 2025

Matrix-based DNA Extraction for Targeted Next-Generation Sequencing on Decontaminated Sputum Samples

Matrix-based DNA Extraction for Targeted Next-Generation Sequencing on Decontaminated Sputum Samples

Published on: June 6, 2025

Optimized Method for Mycobacteria DNA Extraction from Sputum for Isothermal Amplification.

Byoung-Sun Mun¹, Jung Yoon², Soo-Young Yoon¹

¹Department of Laboratory Medicine, Korea University College of Medicine, Seoul, Korea.

Annals of Clinical and Laboratory Science

|July 12, 2023

Summary

A new, simple DNA extraction method using thermal lysis at 65°C with guanidinium thiocyanate (GuSCN) and spin column purification is effective for mycobacterial detection. This robust protocol is suitable for downstream isothermal amplification techniques.

Keywords:

DNA extraction Mycobacterium tuberculosis nontuberculous mycobacteria sputum

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Last Updated: Jul 23, 2025

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Identification of Mycobacterium Species by DNA Microarray Chip Method

Identification of Mycobacterium Species by DNA Microarray Chip Method

Published on: June 24, 2025

Area of Science:

Molecular Biology
Microbiology
Biotechnology

Background:

Mycobacterial DNA extraction from sputum is complex, often requiring multiple centrifugation steps and manual handling.
Existing methods pose challenges for integration with rapid isothermal amplification techniques.

Purpose of the Study:

To establish a simple, robust mycobacterial DNA extraction protocol from sputum samples.
To develop a method suitable for downstream isothermal amplification.

Main Methods:

Evaluated DNA extraction efficiency using spiked sputum samples with *Mycovacterium gordonae* and *M. bovis* bacillus Calmette-Guerin (BCG).
Compared three mycobacterial lysis methods: sonication and heating at 100°C or 65°C.
Assessed optimal DNA adsorption for spin column purification.

Main Results:

Thermal lysis at 65°C for 30 minutes was selected for its ease of use, showing minimal difference in cycle threshold values between tested mycobacteria.
A one-step protocol involving guanidinium thiocyanate (GuSCN) addition before thermal lysis and spin column purification yielded efficient DNA extraction.

Conclusions:

The developed DNA extraction protocol, utilizing thermal lysis at 65°C with GuSCN and spin column purification, is simple, fast (<50 min), and robust.
This protocol is well-suited for downstream isothermal amplification, improving mycobacterial detection workflows.