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Comparison of eleven methods for genomic DNA extraction suitable for large-scale whole-genome genotyping and long-term DNA banking using blood samples

Psifidi, A; Dovas, C I; Bramis, G; Lazou, T; Russel, C L; Arsenos, G; Banos, G

Authors

A Psifidi

C I Dovas

G Bramis

T Lazou

C L Russel

G Arsenos

G Banos



Abstract

Over the recent years, next generation sequencing and microarray technologies have revolutionized scientific research with their applications to high-throughput analysis of biological systems. Isolation of high quantities of pure, intact, double stranded, highly concentrated, not contaminated genomic DNA is prerequisite for successful and reliable large scale genotyping analysis. High quantities of pure DNA are also required for the creation of DNA-banks. In the present study, eleven different DNA extraction procedures, including phenol-chloroform, silica and magnetic beads based extractions, were examined to ascertain their relative effectiveness for extracting DNA from ovine blood samples. The quality and quantity of the differentially extracted DNA was subsequently assessed by spectrophotometric measurements, Qubit measurements, real-time PCR amplifications and gel electrophoresis. Processing time, intensity of labor and cost for each method were also evaluated. Results revealed significant differences among the eleven procedures and only four of the methods yielded satisfactory outputs. These four methods, comprising three modified silica based commercial kits (Modified Blood, Modified Tissue, Modified Dx kits) and an in-house developed magnetic beads based protocol, were most appropriate for extracting high quality and quantity DNA suitable for large-scale microarray genotyping and also for long-term DNA storage as demonstrated by their successful application to 600 individuals.

Citation

Psifidi, A., Dovas, C. I., Bramis, G., Lazou, T., Russel, C. L., Arsenos, G., & Banos, G. (2015). Comparison of eleven methods for genomic DNA extraction suitable for large-scale whole-genome genotyping and long-term DNA banking using blood samples. PLoS ONE, 10(1), e0115960. https://doi.org/10.1371/journal.pone.0115960

Journal Article Type Article
Acceptance Date Nov 28, 2014
Publication Date Jan 30, 2015
Deposit Date Feb 22, 2018
Publicly Available Date Mar 29, 2024
Journal PLoS One
Publisher Public Library of Science
Peer Reviewed Peer Reviewed
Volume 10
Issue 1
Pages e0115960
DOI https://doi.org/10.1371/journal.pone.0115960
Public URL https://rvc-repository.worktribe.com/output/1402090

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