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 Metrics

CrossRef Citations:
0
Total Statistics:

Full-Text HTML Views: 171
Abstract HTML Views: 237
PDF Downloads: 118
Total Views/Downloads: 528
Unique Statistics:

Full-Text HTML Views: 96
Abstract HTML Views: 170
PDF Downloads: 97
Total Views/Downloads: 365



© 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.