The Integrity and Yield of Genomic DNA Isolated from Whole Blood Following Long-Term Storage at −30°C

全血 DNA 基因组DNA DNA提取 琼脂糖凝胶电泳 色谱法 生物 琼脂糖 分子生物学 化学 聚合酶链反应 遗传学 基因 免疫学
作者
Wenlong Carl Chen,Robyn Kerr,Andrew K. May,Babongile C. Ndlovu,Anelisa Sobalisa,Sanelisiwe T. Duze,Lavania Joseph,Christopher G. Mathew,Chantal Babb de Villiers
出处
期刊:Biopreservation and Biobanking [Mary Ann Liebert]
卷期号:16 (2): 106-113 被引量:30
标识
DOI:10.1089/bio.2017.0050
摘要

Long-term storage of whole blood can affect the integrity of DNA if it is not done under optimal conditions. The aim of this study was to determine whether long-term storage (2-19 years) of whole blood samples at -30°C had a negative effect on the quality or quantity of genomic DNA that could be recovered at extraction. Genomic DNA was isolated from 2758 whole blood samples collected in 4 mL EDTA vacutainers from 1997 to 2012. DNA was extracted using the Qiagen® FlexiGene® DNA kit. The average storage duration at -30°C was 12 years. The quality and quantity of the isolated DNA were assessed using spectrophotometry (NanoDrop™), a fluorometric assay for double-stranded DNA (Qubit™), and agarose gel electrophoresis. The mean DNA yield per sample was found to be 114 μg from whole blood volumes that ranged from 0.5 to 4 mL. The mean A260/280 ratio and median A260/280 ratios were both 1.8. No correlation was found between the duration of storage and the total yield or the quality of DNA extracted. These data suggest that high-quality DNA can be extracted from whole blood samples that are stored at -30°C for up to 19 years.

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