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Kopfnagel, V.* ; Klopp, N.* ; Bernemann, I.* ; Nizhegorodtseva, N.* ; Wilson, R. ; Gronauer, R.* ; Seifert, M.* ; Illig, T.*

Effects of repeated freeze and thaw cycles on the genome-wide DNA methylation profile of isolated genomic DNA.

Biopreserv. Biobank. 22, 110-114 (2024)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
The characterization of DNA methylation patterns to identify epigenetic markers for complex human diseases is an important and rapidly evolving part in biomedical research. DNA samples collected and stored in clinical biobanks over the past years are an important source for future epigenetic studies. Isolated gDNA is considered stable when stored at low temperatures for several years. However, the effect of multiple use and the associated repeated thawing of long-term stored DNA samples on DNA methylation patterns has not yet been investigated. In this study, we examined the influence of up to 10 freeze and thaw cycles on global DNA methylation by comparing genome-wide methylation profiles. DNA samples from 19 healthy volunteers were either frozen at -80°C or subjected to up to 10 freeze and thaw cycles. Genome-wide DNA methylation was analyzed after 0, 1, 3, 5, or 10 thaw cycles using the Illumina Infinium MethylationEPIC BeadChip. Evaluation of the global DNA methylation profile by beta-value density plots and multidimensional scaling plots revealed an expected clear participant-dependent variability, but a very low variability depending on the freeze and thaw cycles. In accordance, no significant difference in any of the methylated cytosine/guanine sites studied could be detected in the performed statistical analyses. Our results suggest that long-term frozen DNA samples are still suitable for epigenetic studies after multiple thaw cycles.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Dna Sample Quality ; Biobanking ; Genomic Dna Storage; Peripheral-blood
ISSN (print) / ISBN 1947-5535
e-ISSN 1947-5543
Quellenangaben Volume: 22, Issue: 2, Pages: 110-114 Article Number: , Supplement: ,
Publisher Mary Ann Liebert
Publishing Place New Rochelle, NY
Non-patent literature Publications
Reviewing status Peer reviewed