gDNA extraction yield and methylation status of blood samples are affected by long-term storage conditions.

Schröder, Charlotte; Steimer, Werner

doi:10.1371/journal.pone.0192414

journal article

Title:: gDNA extraction yield and methylation status of blood samples are affected by long-term storage conditions.
Document type:: Journal Article; Research Support, Non-U.S. Gov't; Journal Article; Research Support, Non-U.S. Gov't
Author(s):: Schröder, Charlotte; Steimer, Werner
Abstract:: Epigenetics is believed to provide great chances for a better understanding of the development and treatment of many diseases where the analysis of genomic DNA has so far failed to provide conclusive answers. Methylcytosine is a frequently used quantitative marker of epigenetic studies. Since immediate analysis of sampled material is in most cases not possible, storage time and conditions are critical aspects regarding the quality of genomic DNA and reliability of analysis. Blood is frequently used for such analyses. We, therefore, collected blood samples of ten volunteers and stored them under various conditions for ten months: -70°C, -20°C, 2-8°C and room temperature. An additional aliquot was frozen at -70°C and thawed once a week at room temperature. We then compared the DNA extraction yields and methylation status in relation to storage time and conditions. We found significantly lower DNA extraction yields (up to -97.45%; p <= 0.001) as well as significantly higher methylation levels after ten months of storage (up to +42.0%; p <= 0.001). These results suggest that storage time has an important influence on DNA analyses of blood samples for all storage conditions. This might be due to differences in stability of methylated and non-methylated DNA. Our study indicates that storage conditions and time may be a critical factor for epigenetic methylation studies and require rigorous validation. For reliable analyses we, therefore, recommend to perform epigenetic analysis directly after sample collection.
Journal title abbreviation:: PLoS ONE
Year:: 2018
Journal volume:: 13
Journal issue:: 2
Pages contribution:: e0192414
Language:: eng
Fulltext / DOI:: doi:10.1371/journal.pone.0192414
Pubmed ID:: http://view.ncbi.nlm.nih.gov/pubmed/29415017
Print-ISSN:: 1932-6203
TUM Institution:: Institut für Klinische Chemie und Pathobiochemie
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mediaTUM Gesamtbestand Einrichtungen Schools TUM School of Medicine and Health Departments Clinical Medicine Institut für Klinische Chemie und Pathobiochemie (Prof. Ruland)2018

mediaTUM Gesamtbestand Hochschulbibliographie 2018 Fakultäten Medizin Institut für Klinische Chemie und Pathobiochemie