DNA extraction from clotted blood in genotyping quality

被引：5
作者：
Stanzick, Kira J. [1 ]
Simon, Josef [1 ]
Zimmermann, Martina E. [1 ]
Schachtner, Michael [2 ]
Peterhoff, David [2 ,3 ]
Niller, Hans-Helmut [2 ]
ueberla, Klaus [4 ]
Wagner, Ralf [2 ,3 ]
Heid, Iris M. [1 ]
Stark, Klaus J. [1 ]
机构：
[1] Univ Regensburg, Dept Genet Epidemiol, Franz Josef Str Allee 11, D-93053 Regensburg, Germany
[2] Univ Regensburg, Mol Microbiol Virol, Inst Med Microbiol & Hyg, Franz Josef Str Allee 11, D-93053 Regensburg, Germany
[3] Univ Hosp Regensburg, Inst Clin Microbiol & Hyg, Franz Josef Str Allee 11, D-93053 Regensburg, Germany
[4] Friedrich Alexander Univ Erlangen Nurnberg, Univ Hosp Erlangen, Inst Clin & Mol Virol, Schlossgarten 4, D-91054 Erlangen, Germany
来源：
BIOTECHNIQUES | 2023年 / 74卷 / 01期
关键词：
anticoagulated blood; AugUR; clotted blood; DNA concentration; DNA extraction; DNA purity; DNA yield; frozen samples; genotyping; TiKoCo; YIELD;
D O I：
10.2144/btn-2022-0061
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
引用
收藏
页码：23 / 29
页数：7
相关论文
共 50 条
[31]   High-throughput DNA Extraction and Genotyping of 3dpf Zebrafish Larvae by Fin Clipping [J].
Kosuta, Ceres ;
Daniel, Kate ;
Johnstone, Devon L. ;
Mongeon, Kevin ;
Ban, Kevin ;
LeBlanc, Sophie ;
MacLeod, Stuart ;
Et-Tahiry, Karim ;
Ekker, Marc ;
MacKenzie, Alex ;
Pena, Izabella .
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2018, (136)
[32]   Saliva DNA quality and genotyping efficiency in a predominantly elderly population [J].
Harini V. Gudiseva ;
Mark Hansen ;
Linda Gutierrez ;
David W. Collins ;
Jie He ;
Lana D. Verkuil ;
Ian D. Danford ;
Anna Sagaser ;
Anita S. Bowman ;
Rebecca Salowe ;
Prithvi S. Sankar ;
Eydie Miller-Ellis ;
Amanda Lehman ;
Joan M. O’Brien .
BMC Medical Genomics, 9
[33]   Saliva DNA quality and genotyping efficiency in a predominantly elderly population [J].
Gudiseva, Harini V. ;
Hansen, Mark ;
Gutierrez, Linda ;
Collins, David W. ;
He, Jie ;
Verkuil, Lana D. ;
Danford, Ian D. ;
Sagaser, Anna ;
Bowman, Anita S. ;
Salowe, Rebecca ;
Sankar, Prithvi S. ;
Miller-Ellis, Eydie ;
Lehman, Amanda ;
O'Brien, Joan M. .
BMC MEDICAL GENOMICS, 2016, 9
[34]   Low-Cost and Rapid Method of DNA Extraction from Scaled Fish Blood and Skin Mucus [J].
Gui, Lang ;
Li, Xinyu ;
Lin, Shentao ;
Zhao, Yun ;
Lin, Peiyao ;
Wang, Bingqi ;
Tang, Rongkang ;
Guo, Jing ;
Zu, Yao ;
Zhou, Yan ;
Li, Mingyou .
VIRUSES-BASEL, 2022, 14 (04)
[35]   Extraction of high quality DNA from biological materials and calcified tissues [J].
Stray, James ;
Holt, Allison ;
Brevnov, Maxim ;
Calandro, Lisa M. ;
Furtado, Manohar R. ;
Shewale, Jaiprakash G. .
FORENSIC SCIENCE INTERNATIONAL GENETICS SUPPLEMENT SERIES, 2009, 2 (01) :159-160
[36]   EXTRACTION OF HIGH-QUALITY DNA FROM BLOODSTAINS USING DIATOMS [J].
GUNTHER, S ;
HEROLD, J ;
PATZELT, D .
INTERNATIONAL JOURNAL OF LEGAL MEDICINE, 1995, 108 (03) :154-156
[37]   A protocol for high-quality genomic DNA extraction from legumes [J].
Agbagwa, I. O. ;
Datta, S. ;
Patil, P. G. ;
Singh, P. ;
Nadarajan, N. .
GENETICS AND MOLECULAR RESEARCH, 2012, 11 (04) :4632-4639
[38]   Quantity and Quality Assessment of DNA Extracted from Saliva and Blood [J].
Looi, Mee-Lee ;
Zakaria, Hasbullani ;
Osman, Junaida ;
Jamal, Rahman .
CLINICAL LABORATORY, 2012, 58 (3-4) :307-312
[39]   Higher DNA Yield for Epidemiological Studies: A Better Method for DNA Extraction from Blood Clot [J].
Zhou, Guangdi ;
Du, Yatao ;
Wu, Yuhang ;
Wang, Weiye C. .
GENETIC TESTING AND MOLECULAR BIOMARKERS, 2019, 23 (01) :66-72
[40]   Comparison and Optimization of DNA Extraction Methods for Human DNA from Dried Blood Spot Samples [J].
Van Biesen, Natalja ;
Cools, Piet ;
Meyers, Eline .
PEDIATRIC REPORTS, 2025, 17 (02)
12345