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使用盐析法和自动分离法(MagNA系统)进行DNA分离的比较。

Comparison of DNA isolation using salting-out procedure and automated isolation (MagNA system).

作者信息

Kalousová Marta, Levová Kateřina, Kuběna Aleš A, Jáchymová Marie, Franková Věra, Zima Tomáš

机构信息

a Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine , Charles University and General University Hospital , Prague , Czech Republic.

b Institute of Inherited Metabolic Disorders, First Faculty of Medicine , Charles University and General University Hospital , Prague , Czech Republic.

出版信息

Prep Biochem Biotechnol. 2017 Aug 9;47(7):703-708. doi: 10.1080/10826068.2017.1303613. Epub 2017 Mar 9.

Abstract

Isolation of genomic DNA is a key step in genetic analysis. The aim of the study was to evaluate the suitability of isolation of DNA from peripheral blood with manual salting-out procedure and automated MagNA system under specific conditions. The impact of storage conditions, type of material (whole blood or blood cells), and method used for DNA extraction were evaluated in terms of DNA yield, its purity, and integrity. Fresh material, and material stored at 2-8°C for 1-4 weeks and frozen at -80°C were tested. For fresh samples, salting-out method gives higher yield than MagNA, irrespectively, on material used. Neither the yield of salting-out method nor its purity decreases during the storage of the samples in the fridge (2-8°C) during 4 weeks. Concerning MagNA, storage of blood cells in the fridge decreases the yield of DNA as well as its purity. For frozen samples, for whole blood, MagNA gives better results while for blood cells, salting-out method seems to be better. For fresh samples, salting-out method is the preferred one, and both whole blood and blood cells can be used. For frozen samples, the preferred method depends on the material.

摘要

基因组DNA的分离是基因分析中的关键步骤。本研究的目的是评估在特定条件下,采用手工盐析法和自动化MagNA系统从外周血中分离DNA的适用性。从DNA产量、纯度和完整性方面评估了储存条件、材料类型(全血或血细胞)以及用于DNA提取的方法的影响。测试了新鲜材料、在2-8°C下储存1-4周以及在-80°C下冷冻的材料。对于新鲜样品,无论使用何种材料,盐析法的产量均高于MagNA法。在冰箱(2-8°C)中储存4周期间,盐析法的产量及其纯度均未降低。关于MagNA法,将血细胞储存在冰箱中会降低DNA产量及其纯度。对于冷冻样品,对于全血,MagNA法效果更好,而对于血细胞,盐析法似乎更好。对于新鲜样品,盐析法是首选方法,全血和血细胞均可使用。对于冷冻样品,首选方法取决于材料。

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