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全血样本手动与自动核酸提取方法的比较。

Comparison of manual and automated nucleic acid extraction from whole-blood samples.

作者信息

Riemann Kathrin, Adamzik Michael, Frauenrath Stefan, Egensperger Rupert, Schmid Kurt W, Brockmeyer Norbert H, Siffert Winfried

机构信息

Institute of Pharmacogenetics, University Hospital Essen, Essen, Germany.

出版信息

J Clin Lab Anal. 2007;21(4):244-8. doi: 10.1002/jcla.20174.

Abstract

Nucleic acid extraction and purification from whole blood is a routine application in many laboratories. Automation of this procedure promises standardized sample treatment, a low error rate, and avoidance of contamination. The performance of the BioRobot M48 (Qiagen) and the manual QIAmp DNA Blood Mini Kit (Qiagen) was compared for the extraction of DNA from whole blood. The concentration and purity of the extracted DNAs were determined by spectrophotometry. Analytical sensitivity was assessed by common PCR and genotyping techniques. The quantity and quality of the generated DNAs were slightly higher using the manual extraction method. The results of downstream applications were comparable to each other. Amplification of high-molecular-weight PCR fragments, genotyping by restriction digest, and pyrosequencing were successful for all samples. No cross-contamination could be detected. While automated DNA extraction requires significantly less hands-on time, it is slightly more expensive than the manual extraction method.

摘要

从全血中提取和纯化核酸是许多实验室的常规操作。该过程的自动化有望实现标准化的样品处理、低错误率并避免污染。比较了BioRobot M48(Qiagen公司)和手动QIAmp DNA Blood Mini试剂盒(Qiagen公司)从全血中提取DNA的性能。通过分光光度法测定提取的DNA的浓度和纯度。通过常规PCR和基因分型技术评估分析灵敏度。使用手动提取方法生成的DNA的数量和质量略高。下游应用的结果彼此相当。所有样品的高分子量PCR片段扩增、限制性消化基因分型和焦磷酸测序均成功。未检测到交叉污染。虽然自动化DNA提取所需的实际操作时间明显更少,但它比手动提取方法略贵。

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