Comparison of manual and automated nucleic acid extraction from whole-blood samples.
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.
Riemann
Kathrin
K
Institute of Pharmacogenetics, University Hospital Essen, Essen, Germany. kathrin.riemann@uk-essen.de
Adamzik
Michael
M
Frauenrath
Stefan
S
Egensperger
Rupert
R
Schmid
Kurt W
KW
Brockmeyer
Norbert H
NH
Siffert
Winfried
W
D001769:Blood / Q000821:virology*; D019411:Clinical Laboratory Techniques / Q000191:economics / Q000295:instrumentation* / Q000379:methods*; D004279:DNA, Viral / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D012371:Robotics
D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic