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*, 1
1 Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich Heine University, Düsseldorf, Germany
2 Department of Urology, University Hospital Zürich, Switzerland

Article Information


Identifiers and Pagination:

Year: 2011
Volume: 4
First Page: 15
Last Page: 17
Publisher ID: TOBIOMJ-4-15
DOI: 10.2174/1875318301104010015

Article History:

Received Date: 31/08/2011
Revision Received Date: 25/09/2011
Acceptance Date: 08/10/2011
Electronic publication date: 2/12/2011
Collection year: 2011

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Creative Commons License
© 2011 Beermann et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich Heine University, Düsseldorf, Germany; Tel: +49-(0)211-81-16905; Fax: +49-(0)-211-81-04543; E-mail: santourlidis@itz.uni-duesseldorf.de
# contributed equally.


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.