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Updated: Jul 10, 2026
Filtration Isolation of Nucleic Acids: A Simple and Rapid DNA Extraction Method
Published on: August 6, 2016
Preparation of milk samples for PCR analysis using a rapid filtration technique
1Department of Life Sciences, National Central University, Jung-da Road, Jhong-li City, Taiwan. jyewu@cc.ncu.edu.tw
Aims:
To investigate the usefulness of a straightforward filtration method for the isolation of Escherichia coli O157:H7 contaminants from milk for PCR detection.
Methods And Results:
Escherichia coli O157:H7 is grown in milk and enriched in Luria-Bertani (LB) medium. Samples are filtered through a 0.45-microm pore membrane. The membrane is immersed in 200-microl lysis buffer and incubated at 95 degrees C for 10 min to release bacterial DNA for subsequent PCR detection. Under current conditions, the overall duration from filtration to PCR-ready DNA generation is <20 min, and the detection level for PCR was as low as 10 CFU of bacteria in 1 ml of milk.
Conclusion:
Bacterial contaminants of milk can be concentrated and isolated by a simple, one-step filtration and their DNA can be released for subsequent PCR detection by heating the filter membrane at 95 degrees C for 10 min.
Significance And Impact Of The Study:
The simplicity of this method allows inexpensive, high throughput automation that meets the demands of modern food hygiene monitoring.
