Stanzick, Kira J. and Simon, Josef and Zimmermann, Martina E. and Schachtner, Michael and Peterhoff, David and Niller, Hans-Helmut and ueberla, Klaus and Wagner, Ralf and Heid, Iris M. and Stark, Klaus J. (2023) DNA extraction from clotted blood in genotyping quality. BIOTECHNIQUES, 74 (1). pp. 23-29. ISSN 0736-6205, 1940-9818
Full text not available from this repository. (Request a copy)Abstract
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 mu g 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). METHOD SUMMARYThis study describes DNA extraction from frozen clotted blood using QIAGEN Clotspin Baskets and the Gentra Puregene Blood Kit. The concentration, yield per milliliter of blood, purity and integrity of the DNA were further confirmed via spectrophotometer and gel electrophoresis. In addition, performance for genotyping on the Illumina Global Screening Array was analyzed. Comparisons with DNA extracted from anticoagulated blood were performed.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | YIELD; anticoagulated blood; AugUR; clotted blood; DNA concentration; DNA extraction; DNA purity; DNA yield; frozen samples; genotyping; TiKoCo |
| Subjects: | 500 Science > 570 Life sciences 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene Medicine > Institut für Epidemiologie und Präventivmedizin > Lehrstuhl für Genetische Epidemiologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 10 Mar 2024 10:33 |
| Last Modified: | 10 Mar 2024 10:33 |
| URI: | https://pred.uni-regensburg.de/id/eprint/59538 |
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