DNA extraction from clotted blood in genotyping quality
Kira J Stanzick,
Josef Simon,
Martina E Zimmermann,
Michael Schachtner,
David Peterhoff,
Hans-Helmut Niller,
Klaus Überla,
Ralf Wagner,
Iris M Heid,
Klaus J Stark
Affiliations
Kira J Stanzick
1Department of Genetic Epidemiology, University of Regensburg, Franz-Josef-Strauß-Allee 11, Regensburg, 93053, Germany
Josef Simon
1Department of Genetic Epidemiology, University of Regensburg, Franz-Josef-Strauß-Allee 11, Regensburg, 93053, Germany
Martina E Zimmermann
1Department of Genetic Epidemiology, University of Regensburg, Franz-Josef-Strauß-Allee 11, Regensburg, 93053, Germany
Michael Schachtner
2Institute of Medical Microbiology & Hygiene, Molecular Microbiology (Virology), University of Regensburg, Franz-Josef-Strauß-Allee 11, Regensburg, 93053, Germany
David Peterhoff
2Institute of Medical Microbiology & Hygiene, Molecular Microbiology (Virology), University of Regensburg, Franz-Josef-Strauß-Allee 11, Regensburg, 93053, Germany
Hans-Helmut Niller
2Institute of Medical Microbiology & Hygiene, Molecular Microbiology (Virology), University of Regensburg, Franz-Josef-Strauß-Allee 11, Regensburg, 93053, Germany
Klaus Überla
4Institute of Clinical & Molecular Virology, University Hospital Erlangen, Friedrich-Alexander Universität Erlangen-Nürnberg, Schlossgarten 4, Erlangen, 91054, Germany
Ralf Wagner
2Institute of Medical Microbiology & Hygiene, Molecular Microbiology (Virology), University of Regensburg, Franz-Josef-Strauß-Allee 11, Regensburg, 93053, Germany
Iris M Heid
1Department of Genetic Epidemiology, University of Regensburg, Franz-Josef-Strauß-Allee 11, Regensburg, 93053, Germany
Klaus J Stark
1Department of Genetic Epidemiology, University of Regensburg, Franz-Josef-Strauß-Allee 11, Regensburg, 93053, Germany
DNA extraction from frozen blood clots is challenging. Here, the authors applied QIAGEN Clotspin Baskets and the Gentra Puregene Blood Kit for DNA extraction to cellular fraction of 5.5 ml whole blood without anticoagulating additives. The amount and quality of extracted DNA were assessed via spectrophotometer and gel electrophoresis. Results from array-based genotyping were analyzed. All steps were compared with DNA isolated from anticoagulated blood samples from a separate study. The quality and concentration of DNA extracted from clotted blood were comparable to those of DNA extracted from anticoagulated blood. DNA yield was on average 27 μg per ml clotted blood, with an average purity of 1.87 (A260/A280). Genotyping quality was similar for both DNA sources (call rate: 99.56% from clotted vs 99.49% from anticoagulated blood).
