Simple and cost-effective laboratory methods to evaluate and validate cell-free DNA isolation
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Updated: Feb 3, 2026

Analysis of Fecal Microbiota Dynamics in Lupus-Prone Mice Using a Simple, Cost-Effective DNA Isolation Method
Published on: May 2, 2022
Afsaneh Mojtabanezhad Shariatpanahi1, Parisa Rokni1,2, Elaheh Shahabi1,3
1Cancer Genetics Research Unit, Reza Radiotherapy and Oncology Center, Mashhad, Iran.
Objective:
In the present study, we investigated different simple and cost effective methods to evaluate and validate cell free DNA (cfDNA) isolation. The ability of the QIAamp DNA Blood Mini Kit method to extract cfDNA was assessed by several approaches, including purification of endogenous cfDNA and exogenous spike-in control material, prior to plasma extraction, and followed by quantitative-PCR.
Results:
Using QIAamp DNA Blood Mini kit, nearly 27% (380 bp) to 35% (173 bp) cfDNA was recovered with a higher recovery of smaller size cfDNA (173 bp) in comparison to larger ones (380 bp). These simple laboratory methods can be used to assess the efficiency of any cfDNA isolation method.
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In the visual method, metal-ion indicators (metallochromic dyes), which have distinct colors in their free and complex forms, are added to the mixture to signal the titration's end point. They form stable complexes with metal ions, but these complexes are weaker than the corresponding metal–EDTA complexes. As a...
Data Validation
Key parameters for method validation include:

