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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...

DNA Isolation

DNA Isolation

DNA from cells is required for many biotechnology and research applications, such as molecular cloning. To remove and purify DNA from cells, researchers use various methods of DNA extraction. While the specifics of different protocols may vary, some general concepts underlie the process of DNA extraction.

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Updated: Dec 15, 2025

A Method for Targeted 16S Sequencing of Human Milk Samples

A Method for Targeted 16S Sequencing of Human Milk Samples

Published on: March 23, 2018

DNA extraction method influences human milk bacterial profiles.

A S Cheema¹, L F Stinson¹, C T Lai¹

¹School of Molecular Sciences, The University of Western Australia, Perth, WA, Australia.

Journal of Applied Microbiology

|July 13, 2020

Summary

This summary is machine-generated.

The Qiagen MagAttract Microbial DNA Isolation Kit (QM) is the most effective for extracting bacterial DNA from human milk. Method choice significantly impacts DNA recovery and bacterial community profiles.

Keywords:

DNA extraction contamination full-length 16S rRNA gene sequencing human milk microbiome

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Area of Science:

Microbiology
Molecular Biology
Genomics

Background:

Accurate bacterial DNA extraction from human milk is crucial for understanding infant gut microbiome development.
Existing DNA extraction methods vary in efficiency and may introduce bias.

Purpose of the Study:

To compare the efficacy of four distinct DNA extraction methods for bacterial DNA recovery from human milk.
To identify the optimal method for reliable bacterial DNA isolation and subsequent 16S rRNA gene sequencing.

Main Methods:

Four DNA extraction methods were evaluated: Qiagen MagAttract Microbial DNA Isolation Kit (QM), Norgen Milk Bacterial DNA Isolation Kit (NM), Qiagen MagAttract Microbiome DNA/RNA Isolation Kit (MM), and TRIzol LS Reagent (LS).
Extracted DNA was analyzed via full-length 16S rRNA gene sequencing.
Bacterial DNA yield and contamination levels were assessed.

Main Results:

Kits MM and method LS failed to extract detectable DNA from 9 out of 11 samples.
Kits NM and QM successfully recovered bacterial DNA from all samples, with mean yields of 0.68 ng/μl and 0.55 ng/μl, respectively.
The dominant bacterial species identified by kits NM and QM included Staphylococcus epidermidis, Streptococcus vestibularis, Propionibacterium acnes, Veillonella dispar, and Rothia mucilaginosa. Contamination varied, with QM showing one species and NM showing six in negative controls.

Conclusions:

The Qiagen MagAttract Microbial DNA Isolation Kit (QM) is the most suitable method for bacterial DNA extraction from human milk among those tested.
The selection of DNA extraction method critically influences the efficiency of bacterial DNA recovery and the resulting bacterial community composition profiles.