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
Identifiers and Pagination:Year: 2011
First Page: 15
Last Page: 17
Publisher Id: TOBIOMJ-4-15
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
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.
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.