Human DNA extraction from whole saliva that was fresh or stored for 3, 6 or 12 months using five different protocols
Identifieur interne : 000219 ( Main/Exploration ); précédent : 000218; suivant : 000220Human DNA extraction from whole saliva that was fresh or stored for 3, 6 or 12 months using five different protocols
Auteurs : Thais Francini Garbieri [Brésil] ; Daniel Thomas Brozoski [Brésil] ; Thiago José Dionísio [Brésil] ; Carlos Ferreira Santos [Brésil] ; Lucimara Teixeira Das Neves [Brésil]Source :
- Journal of Applied Oral Science [ 1678-7757 ] ; 2017.
Abstract
Saliva when compared to blood collection has the following advantages: it requires no specialized personnel for collection, allows for remote collection by the patient, is painless, well accepted by participants, has decreased risks of disease transmission, does not clot, can be frozen before DNA extraction and possibly has a longer storage time.
and
This study aimed to compare the quantity and quality of human DNA extracted from saliva that was fresh or frozen for three, six and twelve months using five different DNA extraction protocols: protocol 1 – Oragene™ commercial kit, protocol 2 – QIAamp DNA mini kit, protocol 3 – DNA extraction using ammonium acetate, protocol 4 – Instagene™ Matrix and protocol 5 – Instagene™ Matrix diluted 1:1 using proteinase K and 1% SDS. Briefly, DNA was analyzed using spectrophotometry, electrophoresis and PCR.
Results indicated that time spent in storage typically decreased the DNA quantity with the exception of protocol 1. The purity of DNA was generally not affected by storage times for the commercial based protocols, while the purity of the DNA samples extracted by the noncommercial protocols typically decreased when the saliva was stored longer. Only protocol 1 consistently extracted unfragmented DNA samples. In general, DNA samples extracted through protocols 1, 2, 3 and 4, regardless of storage time, were amplified by human specific primers whereas protocol 5 produced almost no samples that were able to be amplified by human specific primers. Depending on the protocol used, it was possible to extract DNA in high quantities and of good quality using whole saliva, and furthermore, for the purposes of DNA extraction, saliva can be reliably stored for relatively long time periods.
In summary, a complicated picture emerges when taking into account the extracted DNA’s quantity, purity and quality; depending on a given researchers needs, one protocol’s particular strengths and costs might be the deciding factor for its employment.
Url:
DOI: 10.1590/1678-77572016-0046
PubMed: 28403355
PubMed Central: 5393535
Affiliations:
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Le document en format XML
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<front><div type="abstract" xml:lang="en"><title>Abstract</title>
<p>Saliva when compared to blood collection has the following advantages: it requires no specialized personnel for collection, allows for remote collection by the patient, is painless, well accepted by participants, has decreased risks of disease transmission, does not clot, can be frozen before DNA extraction and possibly has a longer storage time. </p>
<sec><title>Objective</title>
<p> and</p>
</sec>
<sec><title>Material and Methods</title>
<p> This study aimed to compare the quantity and quality of human DNA extracted from saliva that was fresh or frozen for three, six and twelve months using five different DNA extraction protocols: protocol 1 – Oragene™ commercial kit, protocol 2 – QIAamp DNA mini kit, protocol 3 – DNA extraction using ammonium acetate, protocol 4 – Instagene™ Matrix and protocol 5 – Instagene™ Matrix diluted 1:1 using proteinase K and 1% SDS. Briefly, DNA was analyzed using spectrophotometry, electrophoresis and PCR.</p>
</sec>
<sec><title>Results</title>
<p> Results indicated that time spent in storage typically decreased the DNA quantity with the exception of protocol 1. The purity of DNA was generally not affected by storage times for the commercial based protocols, while the purity of the DNA samples extracted by the noncommercial protocols typically decreased when the saliva was stored longer. Only protocol 1 consistently extracted unfragmented DNA samples. In general, DNA samples extracted through protocols 1, 2, 3 and 4, regardless of storage time, were amplified by human specific primers whereas protocol 5 produced almost no samples that were able to be amplified by human specific primers. Depending on the protocol used, it was possible to extract DNA in high quantities and of good quality using whole saliva, and furthermore, for the purposes of DNA extraction, saliva can be reliably stored for relatively long time periods.</p>
</sec>
<sec><title>Conclusions</title>
<p> In summary, a complicated picture emerges when taking into account the extracted DNA’s quantity, purity and quality; depending on a given researchers needs, one protocol’s particular strengths and costs might be the deciding factor for its employment.</p>
</sec>
</div>
</front>
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<biblStruct><analytic><author><name sortKey="Schweighardt, Aj" uniqKey="Schweighardt A">AJ Schweighardt</name>
</author>
<author><name sortKey="Tate, Cm" uniqKey="Tate C">CM Tate</name>
</author>
<author><name sortKey="Scott, Ka" uniqKey="Scott K">KA Scott</name>
</author>
<author><name sortKey="Harper, Ka" uniqKey="Harper K">KA Harper</name>
</author>
<author><name sortKey="Robertson, Jm" uniqKey="Robertson J">JM Robertson</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Thiede, C" uniqKey="Thiede C">C Thiede</name>
</author>
<author><name sortKey="Prange Krex, G" uniqKey="Prange Krex G">G Prange-Krex</name>
</author>
<author><name sortKey="Freiberg Richter, J" uniqKey="Freiberg Richter J">J Freiberg-Richter</name>
</author>
<author><name sortKey="Bornh User, M" uniqKey="Bornh User M">M Bornhäuser</name>
</author>
<author><name sortKey="Ehninger, G" uniqKey="Ehninger G">G Ehninger</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Viltrop, T" uniqKey="Viltrop T">T Viltrop</name>
</author>
<author><name sortKey="Krjutskov, K" uniqKey="Krjutskov K">K Krjutskov</name>
</author>
<author><name sortKey="Palta, P" uniqKey="Palta P">P Palta</name>
</author>
<author><name sortKey="Metspalu, A" uniqKey="Metspalu A">A Metspalu</name>
</author>
</analytic>
</biblStruct>
</listBibl>
</div1>
</back>
</TEI>
<affiliations><list><country><li>Brésil</li>
</country>
<region><li>État de São Paulo</li>
</region>
<settlement><li>São Paulo</li>
</settlement>
<orgName><li>Université de São Paulo</li>
</orgName>
</list>
<tree><country name="Brésil"><region name="État de São Paulo"><name sortKey="Garbieri, Thais Francini" sort="Garbieri, Thais Francini" uniqKey="Garbieri T" first="Thais Francini" last="Garbieri">Thais Francini Garbieri</name>
</region>
<name sortKey="Brozoski, Daniel Thomas" sort="Brozoski, Daniel Thomas" uniqKey="Brozoski D" first="Daniel Thomas" last="Brozoski">Daniel Thomas Brozoski</name>
<name sortKey="Dionisio, Thiago Jose" sort="Dionisio, Thiago Jose" uniqKey="Dionisio T" first="Thiago José" last="Dionísio">Thiago José Dionísio</name>
<name sortKey="Garbieri, Thais Francini" sort="Garbieri, Thais Francini" uniqKey="Garbieri T" first="Thais Francini" last="Garbieri">Thais Francini Garbieri</name>
<name sortKey="Neves, Lucimara Teixeira Das" sort="Neves, Lucimara Teixeira Das" uniqKey="Neves L" first="Lucimara Teixeira Das" last="Neves">Lucimara Teixeira Das Neves</name>
<name sortKey="Neves, Lucimara Teixeira Das" sort="Neves, Lucimara Teixeira Das" uniqKey="Neves L" first="Lucimara Teixeira Das" last="Neves">Lucimara Teixeira Das Neves</name>
<name sortKey="Santos, Carlos Ferreira" sort="Santos, Carlos Ferreira" uniqKey="Santos C" first="Carlos Ferreira" last="Santos">Carlos Ferreira Santos</name>
</country>
</tree>
</affiliations>
</record>
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