Collection and Extraction of Saliva DNA for Next Generation Sequencing
Identifieur interne : 000600 ( Pmc/Corpus ); précédent : 000599; suivant : 000601Collection and Extraction of Saliva DNA for Next Generation Sequencing
Auteurs : Michael R. Goode ; Soo Yeon Cheong ; Ning Li ; William C. Ray ; Christopher W. BartlettSource :
- Journal of Visualized Experiments : JoVE [ 1940-087X ] ; 2014.
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.
Url:
DOI: 10.3791/51697
PubMed: 25225892
PubMed Central: 4442313
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PMC:4442313Le document en format XML
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<author><name sortKey="Goode, Michael R" sort="Goode, Michael R" uniqKey="Goode M" first="Michael R." last="Goode">Michael R. Goode</name>
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<author><name sortKey="Cheong, Soo Yeon" sort="Cheong, Soo Yeon" uniqKey="Cheong S" first="Soo Yeon" last="Cheong">Soo Yeon Cheong</name>
<affiliation><nlm:aff id="ID1">Battelle Center for Mathematical Medicine, The Research Institute at Nationwide Children's Hospital</nlm:aff>
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<author><name sortKey="Li, Ning" sort="Li, Ning" uniqKey="Li N" first="Ning" last="Li">Ning Li</name>
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<author><name sortKey="Ray, William C" sort="Ray, William C" uniqKey="Ray W" first="William C." last="Ray">William C. Ray</name>
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<affiliation><nlm:aff id="ID2">The Interdisciplinary Graduate Program in Biophysics, The Ohio State University</nlm:aff>
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<author><name sortKey="Bartlett, Christopher W" sort="Bartlett, Christopher W" uniqKey="Bartlett C" first="Christopher W." last="Bartlett">Christopher W. Bartlett</name>
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<affiliation><nlm:aff id="ID1">Battelle Center for Mathematical Medicine, The Research Institute at Nationwide Children's Hospital</nlm:aff>
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<author><name sortKey="Ray, William C" sort="Ray, William C" uniqKey="Ray W" first="William C." last="Ray">William C. Ray</name>
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<front><div type="abstract" xml:lang="en"><p>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.</p>
</div>
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<pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Vis Exp</journal-id>
<journal-id journal-id-type="iso-abbrev">J Vis Exp</journal-id>
<journal-id journal-id-type="publisher-id">JoVE</journal-id>
<journal-title-group><journal-title>Journal of Visualized Experiments : JoVE</journal-title>
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<issn pub-type="epub">1940-087X</issn>
<publisher><publisher-name>MyJove Corporation</publisher-name>
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<article-id pub-id-type="publisher-id">51697</article-id>
<article-id pub-id-type="doi">10.3791/51697</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Medicine</subject>
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<title-group><article-title>Collection and Extraction of Saliva DNA for Next Generation Sequencing</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Goode</surname>
<given-names>Michael R.</given-names>
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<xref ref-type="aff" rid="ID1"><sup>1</sup>
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<contrib contrib-type="author"><name><surname>Cheong</surname>
<given-names>Soo Yeon</given-names>
</name>
<xref ref-type="aff" rid="ID1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Ning</given-names>
</name>
<xref ref-type="aff" rid="ID1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Ray</surname>
<given-names>William C.</given-names>
</name>
<xref ref-type="aff" rid="ID1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="ID2"><sup>2</sup>
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</contrib>
<contrib contrib-type="author"><name><surname>Bartlett</surname>
<given-names>Christopher W.</given-names>
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<xref ref-type="aff" rid="ID1"><sup>1</sup>
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<xref ref-type="aff" rid="ID3"><sup>3</sup>
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<aff id="ID1"><sup>1</sup>
Battelle Center for Mathematical Medicine, The Research Institute at Nationwide Children's Hospital</aff>
<aff id="ID2"><sup>2</sup>
The Interdisciplinary Graduate Program in Biophysics, The Ohio State University</aff>
<aff id="ID3"><sup>3</sup>
Department of Pediatrics, The Ohio State University</aff>
<author-notes><fn><p>Correspondence to: Christopher W. Bartlett at <email>christopher.bartlett@nationwidechildrens.org</email>
</p>
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</author-notes>
<pub-date pub-type="collection"><year>2014</year>
</pub-date>
<pub-date pub-type="epub"><day>27</day>
<month>8</month>
<year>2014</year>
</pub-date>
<issue>90</issue>
<elocation-id>51697</elocation-id>
<permissions><copyright-statement>Copyright © 2014, Journal of Visualized Experiments</copyright-statement>
<copyright-year>2014</copyright-year>
</permissions>
<abstract><p>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.</p>
</abstract>
<kwd-group kwd-group-type="author-generated"><kwd>Medicine</kwd>
<kwd>Issue 90</kwd>
<kwd>DNA collection</kwd>
<kwd>saliva</kwd>
<kwd>DNA extraction</kwd>
<kwd>Next generation sequencing</kwd>
<kwd>DNA purification</kwd>
<kwd>DNA</kwd>
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