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DNA extraction from clotted blood in genotyping quality

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 spectr...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9887531/
https://www.ncbi.nlm.nih.gov/pubmed/36597257
http://dx.doi.org/10.2144/btn-2022-0061
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author 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
author_facet 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
author_sort Stanzick, Kira J
collection PubMed
description 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).
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spelling pubmed-98875312023-02-03 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 Stark, Klaus J Biotechniques Reports 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). Future Science Ltd 2023-01-04 2023-01 /pmc/articles/PMC9887531/ /pubmed/36597257 http://dx.doi.org/10.2144/btn-2022-0061 Text en © 2023 Klaus J. Stark https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License (https://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Reports
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
title DNA extraction from clotted blood in genotyping quality
title_full DNA extraction from clotted blood in genotyping quality
title_fullStr DNA extraction from clotted blood in genotyping quality
title_full_unstemmed DNA extraction from clotted blood in genotyping quality
title_short DNA extraction from clotted blood in genotyping quality
title_sort dna extraction from clotted blood in genotyping quality
topic Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9887531/
https://www.ncbi.nlm.nih.gov/pubmed/36597257
http://dx.doi.org/10.2144/btn-2022-0061
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