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用于下一代测序的唾液DNA的收集与提取。

Collection and extraction of saliva DNA for next generation sequencing.

作者信息

Goode Michael R, Cheong Soo Yeon, Li Ning, Ray William C, Bartlett Christopher W

机构信息

Battelle Center for Mathematical Medicine, The Research Institute at Nationwide Children's Hospital.

Battelle Center for Mathematical Medicine, The Research Institute at Nationwide Children's Hospital; The Interdisciplinary Graduate Program in Biophysics, The Ohio State University.

出版信息

J Vis Exp. 2014 Aug 27(90):51697. doi: 10.3791/51697.

DOI:10.3791/51697
PMID:25225892
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4442313/
Abstract

The preferred source of DNA in human genetics research is blood, or cell lines derived from blood, as these sources yield large quantities of high quality DNA. However, DNA extraction from saliva can yield high quality DNA with little to no degradation/fragmentation that is suitable for a variety of DNA assays without the expense of a phlebotomist and can even be acquired through the mail. However, at present, no saliva DNA collection/extraction protocols for next generation sequencing have been presented in the literature. This protocol optimizes parameters of saliva collection/storage and DNA extraction to be of sufficient quality and quantity for DNA assays with the highest standards, including microarray genotyping and next generation sequencing.

摘要

在人类遗传学研究中,DNA的首选来源是血液或源自血液的细胞系,因为这些来源能产生大量高质量的DNA。然而,从唾液中提取DNA可以得到高质量的DNA,几乎没有降解/片段化,适用于各种DNA检测,而且无需采血技师,甚至可以通过邮寄获得。然而,目前文献中尚未出现用于下一代测序的唾液DNA收集/提取方案。本方案优化了唾液收集/储存和DNA提取的参数,以获得足够质量和数量的DNA用于最高标准的DNA检测,包括微阵列基因分型和下一代测序。

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本文引用的文献

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Genotyping performance between saliva and blood-derived genomic DNAs on the DMET array: a comparison.唾液和血液来源基因组 DNA 在 DMET 阵列上的基因分型性能比较:一项对比研究。
PLoS One. 2012;7(3):e33968. doi: 10.1371/journal.pone.0033968. Epub 2012 Mar 20.
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A framework for variation discovery and genotyping using next-generation DNA sequencing data.利用下一代 DNA 测序数据进行变异发现和基因分型的框架。
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Increasing genotype-phenotype model determinism: application to bivariate reading/language traits and epistatic interactions in language-impaired families.增强基因型-表型模型的确定性:应用于语言障碍家庭中的双变量阅读/语言特征及上位性相互作用
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5
Saliva-derived DNA performs well in large-scale, high-density single-nucleotide polymorphism microarray studies.唾液来源的 DNA 在大规模、高密度单核苷酸多态性微阵列研究中表现良好。
Cancer Epidemiol Biomarkers Prev. 2010 Mar;19(3):794-8. doi: 10.1158/1055-9965.EPI-09-0812. Epub 2010 Mar 3.
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Cancer Epidemiol Biomarkers Prev. 2007 Oct;16(10):2072-6. doi: 10.1158/1055-9965.EPI-07-0611.
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High microsatellite and SNP genotyping success rates established in a large number of genomic DNA samples extracted from mouth swabs and genotypes.在从口腔拭子提取的大量基因组DNA样本中建立了高微卫星和单核苷酸多态性基因分型成功率及基因型。
Twin Res Hum Genet. 2006 Aug;9(4):501-6. doi: 10.1375/183242706778024973.
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Evaluation of saliva as a source of human DNA for population and association studies.评估唾液作为用于群体和关联研究的人类DNA来源。
Anal Biochem. 2006 Jun 15;353(2):272-7. doi: 10.1016/j.ab.2006.03.021. Epub 2006 Mar 30.
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Collecting saliva by mail for genetic and cotinine analyses in participants recruited through the Internet.通过邮件收集通过互联网招募的参与者的唾液,用于基因和可替宁分析。
Eur J Epidemiol. 2005;20(10):833-8. doi: 10.1007/s10654-005-2148-7.