Methods for Separate Isolation of Cell-Free DNA and Cellular DNA from Urine-Application of Methylation-Specific PCR on both DNA Fractions

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RESEARCH ARTICLE

Methods for Separate Isolation of Cell-Free DNA and Cellular DNA from Urine-Application of Methylation-Specific PCR on both DNA Fractions

Agnes Beermann, 1 Foued Ghanjati, 1 Thomas Hermanns2 Cedric Poyet2 Joana Pereira1 Johannes Fischer1 Peter Wernet1 Simeon Santourlidis, * Open Modal , 1
Authors Info & Affiliations
The Open Biomarkers Journal • 02 Dec 2011 • RESEARCH ARTICLE • DOI: 10.2174/1875318301104010015

Abstract

The analysis of genomic DNA is widely-used for research, forensic and diagnostic purposes. Here we describe reliable methods for isolation of cell-free DNA and cellular DNA from urine. Both DNA fractions are suitable for PCR and Methylation-Specific PCR (MSP) amplification, leading to consistent and reproducible results. A kinetics analysis describes the decline of efficiency of MSP performed with urinary DNA which had been stored at room temperature with and without proteinase K for various time periods.

Keywords: Cell-free DNA, Diagnosis, DNA methylation, Urine.