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Anal Bioanal Chem. 2015 Sep;407(22):6873-8. doi: 10.1007/s00216-015-8846-4. Epub 2015 Jun 28.

Comprehensive evaluation of methods to isolate, quantify, and characterize circulating cell-free DNA from small volumes of plasma.

Author information

1
Laboratory for Epigenetics and Environment, Centre National de Génotypage, CEA-Institut de Génomique, 2 rue Gaston Crémieux, CP 5721, 91000, Evry, France.

Abstract

Circulating cell-free DNA (ccfDNA) has great potential for non-invasive diagnostics, and prediction and monitoring of treatment response, but its amount is usually limited. Therefore, the choice of methods to extract and characterize ccfDNA is crucial. In the current study, we performed the most comprehensive comparison of methods for ccfDNA extraction (11 methods), quantification (3 methods), and estimation of the integrity index (2 methods) from small quantities of different kinds of plasma. The QIAamp® Circulating Nucleic Acid Kit and the Norgen Plasma/Serum Circulating DNA Purification Mini Kit showed the best accuracy and reproducibility, but the Norgen kit allowed to extract a higher amount of ccfDNA. This workflow provides a reliable protocol for the multiple applications of ccfDNA in biomedicine.

PMID:
26123439
DOI:
10.1007/s00216-015-8846-4
[Indexed for MEDLINE]

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