优化从干燥血液样本中提取总 DNA 的方法。

IF 2.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemical Genetics Pub Date : 2024-07-12 DOI:10.1007/s10528-024-10882-7
Jeanne V Samsonova, Nikolay Yu Saushkin, Valery N Voronkova, Yuri A Stolpovsky, Aleksei K Piskunov
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引用次数: 0

摘要

虽然干血斑是一种方便的遗传物质来源,但其 DNA 得率通常低于原生样本。这项研究评估了在玻璃纤维膜和纤维素膜上制备的干血样的 DNA 得率,并调查了得率降低的原因。研究采用旋柱提取法优化了从膜干燥血液样本中提取总 DNA 的过程。结果表明,在水介质中对干燥基质进行初步短期(20 分钟)溶解,然后对洗脱液与膜的混合物进行标准提取,可获得最高的 DNA 得率。玻璃纤维膜的 DNA 产量比原生样本低 40-50%,但平均比传统纤维素膜(滤纸)高两倍。使用干燥样品时 DNA 得率降低的原因是在裂解阶段膜材料截留了部分核酸。
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Optimization of Total DNA Extraction from Dried Blood Samples.

While dried blood spots are a convenient source of genetic material, they are usually associated with a lower DNA yield than from a native sample. The study evaluated the DNA yield from dried blood samples prepared on glass fibre and cellulose membranes and investigated the reasons for the yield reduction. The extraction of total DNA from membrane-dried blood samples was optimized by spin-column extraction method. It was shown that preliminary short-term (20 min) solubilization of a dried matrix in an aqueous medium, followed by standard extraction protocols for the mixture of the eluate with membranes, provides the highest DNA yield. The yield of DNA from a glass fibre membrane was 40-50% lower compared to a native sample, but on average, two times higher than from a conventional cellulose membrane (filter paper). The reduction of DNA yield when using a dried sample was found to be due to partial retention of nucleic acids by the membrane material during the lysis stage.

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来源期刊
Biochemical Genetics
Biochemical Genetics 生物-生化与分子生物学
CiteScore
3.90
自引率
0.00%
发文量
133
审稿时长
4.8 months
期刊介绍: Biochemical Genetics welcomes original manuscripts that address and test clear scientific hypotheses, are directed to a broad scientific audience, and clearly contribute to the advancement of the field through the use of sound sampling or experimental design, reliable analytical methodologies and robust statistical analyses. Although studies focusing on particular regions and target organisms are welcome, it is not the journal’s goal to publish essentially descriptive studies that provide results with narrow applicability, or are based on very small samples or pseudoreplication. Rather, Biochemical Genetics welcomes review articles that go beyond summarizing previous publications and create added value through the systematic analysis and critique of the current state of knowledge or by conducting meta-analyses. Methodological articles are also within the scope of Biological Genetics, particularly when new laboratory techniques or computational approaches are fully described and thoroughly compared with the existing benchmark methods. Biochemical Genetics welcomes articles on the following topics: Genomics; Proteomics; Population genetics; Phylogenetics; Metagenomics; Microbial genetics; Genetics and evolution of wild and cultivated plants; Animal genetics and evolution; Human genetics and evolution; Genetic disorders; Genetic markers of diseases; Gene technology and therapy; Experimental and analytical methods; Statistical and computational methods.
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