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胎儿DNA:最佳提取策略

Fetal DNA: strategies for optimal recovery.

作者信息

Legler Tobias J, Heermann Klaus-Hinrich, Liu Zhong, Soussan Aicha Ait, van der Schoot C Ellen

机构信息

Department of Transfusion Medicine, University of Göttingen, Göttingen, Germany.

出版信息

Methods Mol Biol. 2008;444:209-18. doi: 10.1007/978-1-59745-066-9_16.

Abstract

For fetal DNA extraction, in principle each DNA extraction method can be used; however, because most methods have been optimized for genomic DNA from leucocytes, we describe here the methods that have been optimized for the extraction of fetal DNA from maternal plasma and validated for this purpose in our laboratories. The use of the QIAamp DSP Virus kit (QIAGEN), the QIAamp DNA Blood Mini kit (QIAGEN), and the Magna Pure LC (Roche) is based on the kit components provided by the respective companies. However, we noticed that the yield of fetal DNA from maternal plasma can be increased when higher volumes are processed or some slight modifications of the protocols provided by the manufacturer are followed. Here, we also describe an in-house method that allows the specific capture of target molecules in an extremely low volume by using magnetic beads and magnetic tips. This method can be either performed by hand, or it can be adapted to a commercially available pipetting workstation.

摘要

对于胎儿DNA提取,原则上每种DNA提取方法都可以使用;然而,由于大多数方法是针对白细胞中的基因组DNA进行优化的,我们在此描述已针对从母体血浆中提取胎儿DNA进行优化并在我们实验室中为此目的经过验证的方法。使用QIAamp DSP病毒试剂盒(QIAGEN)、QIAamp DNA血液微型试剂盒(QIAGEN)和Magna Pure LC(罗氏)是基于各公司提供的试剂盒组件。然而,我们注意到,当处理更高体积的样本或遵循制造商提供的方案进行一些轻微修改时,母体血浆中胎儿DNA的产量可以提高。在此,我们还描述了一种内部方法,该方法通过使用磁珠和磁棒在极低体积下特异性捕获目标分子。此方法既可以手动进行,也可以适用于市售的移液工作站。

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