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Stanzick, Kira J. ; Simon, Josef ; Zimmermann, Martina E. ; Schachtner, Michael ; Peterhoff, David ; Niller, Hans-Helmut ; Überla, Klaus ; Wagner, Ralf ; Heid, Iris M. ; Stark, Klaus J.

DNA extraction from clotted blood in genotyping quality

Stanzick, Kira J., Simon, Josef, Zimmermann, Martina E. , Schachtner, Michael, Peterhoff, David , Niller, Hans-Helmut, Überla, Klaus, Wagner, Ralf, Heid, Iris M. and Stark, Klaus J. (2023) DNA extraction from clotted blood in genotyping quality. BioTechniques 74 (1).

Date of publication of this fulltext: 18 Jan 2023 13:36
Article
DOI to cite this document: 10.5283/epub.53564


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 ...

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.



Involved Institutions


Details

Item typeArticle
Journal or Publication TitleBioTechniques
Publisher:FUTURE SCI LTD
Place of Publication:LONDON
Volume:74
Number of Issue or Book Chapter:1
Date4 January 2023
InstitutionsMedicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene
Medicine > Institut für Epidemiologie und Präventivmedizin > Lehrstuhl für Genetische Epidemiologie
Identification Number
ValueType
10.2144/btn-2022-0061DOI
KeywordsYIELD; anticoagulated blood; AugUR; clotted blood; DNA concentration; DNA extraction; DNA purity; DNA yield; frozen samples; genotyping; TiKoCo
Dewey Decimal Classification500 Science > 570 Life sciences
600 Technology > 610 Medical sciences Medicine
StatusPublished
RefereedYes, this version has been refereed
Created at the University of RegensburgYes
URN of the UB Regensburgurn:nbn:de:bvb:355-epub-535644
Item ID53564

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