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

被引:56
作者
Riemann, Kathrin
Adamzik, Michael
Frauenrath, Stefan
Egensperger, Rupert
Schmid, Kurt W.
Brockmeyer, Norbert H.
Siffert, Winfried
机构
[1] Univ Hosp Essen, Inst Pharmacogenet, D-45147 Essen, Germany
[2] Univ Hosp Essen, Dept Anaesthesiol, Essen, Germany
[3] Univ Hosp Essen, Inst Pathol & Neuropathol, Essen, Germany
[4] Ruhr Univ Bochum, Hosp Dermatol & Allergy, D-4630 Bochum, Germany
关键词
DNA purification; quality; quantity; roboter; PCR;
D O I
10.1002/jcla.20174
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
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((R)) 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. (c) 2007 Wiley-Liss, Inc.
引用
收藏
页码:244 / 248
页数:5
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