Purification and preconcentration of genomic DNA from whole cell lysates using photoactivated polycarbonate (PPC) microfluidic chips
Identifieur interne : 001625 ( Main/Exploration ); précédent : 001624; suivant : 001626Purification and preconcentration of genomic DNA from whole cell lysates using photoactivated polycarbonate (PPC) microfluidic chips
Auteurs : Małgorzata A. Witek [États-Unis] ; Shawn D. Llopis [États-Unis] ; Abigail Wheatley [États-Unis] ; Robin L. Mccarley [États-Unis] ; Steven A. Soper [États-Unis]Source :
- Nucleic Acids Research [ 0305-1048 ] ; 2006.
Abstract
We discuss the use of a photoactivated polycarbonate (PPC) microfluidic chip for the solid-phase, reversible immobilization (SPRI) and purification of genomic DNA (gDNA) from whole cell lysates. The surface of polycarbonate was activated by UV radiation resulting in a photo-oxidation reaction, which produced a channel surface containing carboxylate groups. The gDNA was selectively captured on this photoactivated surface in an immobilization buffer, which consisted of 3% polyethylene glycol, 0.4 M NaCl and 70% ethanol. The methodology reported herein is similar to conventional SPRI in that surface-confined carboxylate groups are used for the selective immobilization of DNA; however, no magnetic beads or a magnetic field are required. As observed by UV spectroscopy, a load of ∼7.6 ± 1.6 µg/ml of gDNA was immobilized onto the PPC bed. The recovery of DNA following purification was estimated to be 85 ± 5%. The immobilization and purification assay using this PPC microchip could be performed within ∼25 min as follows: (i) DNA immobilization ∼6 min, (ii) chip washout with ethanol 10 min, and (iii) drying and gDNA desorption ∼6 min. The PPC microchip could also be used for subsequent assays with no substantial loss in recovery, no observable carryover and no need for ‘reactivation’ of the PC surface with UV light.
Url:
DOI: 10.1093/nar/gkl146
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 000A25
- to stream Istex, to step Curation: 000A24
- to stream Istex, to step Checkpoint: 001071
- to stream Main, to step Merge: 001633
- to stream Main, to step Curation: 001625
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Purification and preconcentration of genomic DNA from whole cell lysates using photoactivated polycarbonate (PPC) microfluidic chips</title>
<author><name sortKey="Witek, Malgorzata A" sort="Witek, Malgorzata A" uniqKey="Witek M" first="Małgorzata A." last="Witek">Małgorzata A. Witek</name>
</author>
<author><name sortKey="Llopis, Shawn D" sort="Llopis, Shawn D" uniqKey="Llopis S" first="Shawn D." last="Llopis">Shawn D. Llopis</name>
</author>
<author><name sortKey="Wheatley, Abigail" sort="Wheatley, Abigail" uniqKey="Wheatley A" first="Abigail" last="Wheatley">Abigail Wheatley</name>
</author>
<author><name sortKey="Mccarley, Robin L" sort="Mccarley, Robin L" uniqKey="Mccarley R" first="Robin L." last="Mccarley">Robin L. Mccarley</name>
</author>
<author><name sortKey="Soper, Steven A" sort="Soper, Steven A" uniqKey="Soper S" first="Steven A." last="Soper">Steven A. Soper</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">ISTEX</idno>
<idno type="RBID">ISTEX:F204A775A297E50260E05CE15E94036537BAC12F</idno>
<date when="2006" year="2006">2006</date>
<idno type="doi">10.1093/nar/gkl146</idno>
<idno type="url">https://api.istex.fr/document/F204A775A297E50260E05CE15E94036537BAC12F/fulltext/pdf</idno>
<idno type="wicri:Area/Istex/Corpus">000A25</idno>
<idno type="wicri:Area/Istex/Curation">000A24</idno>
<idno type="wicri:Area/Istex/Checkpoint">001071</idno>
<idno type="wicri:doubleKey">0305-1048:2006:Witek M:purification:and:preconcentration</idno>
<idno type="wicri:Area/Main/Merge">001633</idno>
<idno type="wicri:Area/Main/Curation">001625</idno>
<idno type="wicri:Area/Main/Exploration">001625</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Purification and preconcentration of genomic DNA from whole cell lysates using photoactivated polycarbonate (PPC) microfluidic chips</title>
<author><name sortKey="Witek, Malgorzata A" sort="Witek, Malgorzata A" uniqKey="Witek M" first="Małgorzata A." last="Witek">Małgorzata A. Witek</name>
<affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Center for Biomodular Multi-Scale Systems, Louisiana State University Baton Rouge, LA 70803</wicri:regionArea>
<placeName><region type="state">Louisiane</region>
</placeName>
</affiliation>
<affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Chemistry, Louisiana State University 232 Choppin Hall, Baton Rouge, LA 70803</wicri:regionArea>
<placeName><region type="state">Louisiane</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Llopis, Shawn D" sort="Llopis, Shawn D" uniqKey="Llopis S" first="Shawn D." last="Llopis">Shawn D. Llopis</name>
<affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Center for Biomodular Multi-Scale Systems, Louisiana State University Baton Rouge, LA 70803</wicri:regionArea>
<placeName><region type="state">Louisiane</region>
</placeName>
</affiliation>
<affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Chemistry, Louisiana State University 232 Choppin Hall, Baton Rouge, LA 70803</wicri:regionArea>
<placeName><region type="state">Louisiane</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Wheatley, Abigail" sort="Wheatley, Abigail" uniqKey="Wheatley A" first="Abigail" last="Wheatley">Abigail Wheatley</name>
<affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Chemistry, Louisiana State University 232 Choppin Hall, Baton Rouge, LA 70803</wicri:regionArea>
<placeName><region type="state">Louisiane</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Mccarley, Robin L" sort="Mccarley, Robin L" uniqKey="Mccarley R" first="Robin L." last="Mccarley">Robin L. Mccarley</name>
<affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Center for Biomodular Multi-Scale Systems, Louisiana State University Baton Rouge, LA 70803</wicri:regionArea>
<placeName><region type="state">Louisiane</region>
</placeName>
</affiliation>
<affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Chemistry, Louisiana State University 232 Choppin Hall, Baton Rouge, LA 70803</wicri:regionArea>
<placeName><region type="state">Louisiane</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Soper, Steven A" sort="Soper, Steven A" uniqKey="Soper S" first="Steven A." last="Soper">Steven A. Soper</name>
<affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Center for Biomodular Multi-Scale Systems, Louisiana State University Baton Rouge, LA 70803</wicri:regionArea>
<placeName><region type="state">Louisiane</region>
</placeName>
</affiliation>
<affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Chemistry, Louisiana State University 232 Choppin Hall, Baton Rouge, LA 70803</wicri:regionArea>
<placeName><region type="state">Louisiane</region>
</placeName>
</affiliation>
<affiliation wicri:level="1"><country wicri:rule="url">États-Unis</country>
</affiliation>
</author>
</analytic>
<monogr></monogr>
<series><title level="j">Nucleic Acids Research</title>
<title level="j" type="abbrev">Nucl. Acids Res.</title>
<idno type="ISSN">0305-1048</idno>
<idno type="eISSN">1362-4962</idno>
<imprint><publisher>Oxford University Press</publisher>
<date type="published" when="2006">2006</date>
<biblScope unit="volume">34</biblScope>
<biblScope unit="issue">10</biblScope>
<biblScope unit="page" from="74">e74</biblScope>
<biblScope unit="page" to="74">e74</biblScope>
</imprint>
<idno type="ISSN">0305-1048</idno>
</series>
<idno type="istex">F204A775A297E50260E05CE15E94036537BAC12F</idno>
<idno type="DOI">10.1093/nar/gkl146</idno>
<idno type="local">gkl146</idno>
</biblStruct>
</sourceDesc>
<seriesStmt><idno type="ISSN">0305-1048</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass></textClass>
<langUsage><language ident="en">en</language>
</langUsage>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">We discuss the use of a photoactivated polycarbonate (PPC) microfluidic chip for the solid-phase, reversible immobilization (SPRI) and purification of genomic DNA (gDNA) from whole cell lysates. The surface of polycarbonate was activated by UV radiation resulting in a photo-oxidation reaction, which produced a channel surface containing carboxylate groups. The gDNA was selectively captured on this photoactivated surface in an immobilization buffer, which consisted of 3% polyethylene glycol, 0.4 M NaCl and 70% ethanol. The methodology reported herein is similar to conventional SPRI in that surface-confined carboxylate groups are used for the selective immobilization of DNA; however, no magnetic beads or a magnetic field are required. As observed by UV spectroscopy, a load of ∼7.6 ± 1.6 µg/ml of gDNA was immobilized onto the PPC bed. The recovery of DNA following purification was estimated to be 85 ± 5%. The immobilization and purification assay using this PPC microchip could be performed within ∼25 min as follows: (i) DNA immobilization ∼6 min, (ii) chip washout with ethanol 10 min, and (iii) drying and gDNA desorption ∼6 min. The PPC microchip could also be used for subsequent assays with no substantial loss in recovery, no observable carryover and no need for ‘reactivation’ of the PC surface with UV light.</div>
</front>
</TEI>
<affiliations><list><country><li>États-Unis</li>
</country>
<region><li>Louisiane</li>
</region>
</list>
<tree><country name="États-Unis"><region name="Louisiane"><name sortKey="Witek, Malgorzata A" sort="Witek, Malgorzata A" uniqKey="Witek M" first="Małgorzata A." last="Witek">Małgorzata A. Witek</name>
</region>
<name sortKey="Llopis, Shawn D" sort="Llopis, Shawn D" uniqKey="Llopis S" first="Shawn D." last="Llopis">Shawn D. Llopis</name>
<name sortKey="Llopis, Shawn D" sort="Llopis, Shawn D" uniqKey="Llopis S" first="Shawn D." last="Llopis">Shawn D. Llopis</name>
<name sortKey="Mccarley, Robin L" sort="Mccarley, Robin L" uniqKey="Mccarley R" first="Robin L." last="Mccarley">Robin L. Mccarley</name>
<name sortKey="Mccarley, Robin L" sort="Mccarley, Robin L" uniqKey="Mccarley R" first="Robin L." last="Mccarley">Robin L. Mccarley</name>
<name sortKey="Soper, Steven A" sort="Soper, Steven A" uniqKey="Soper S" first="Steven A." last="Soper">Steven A. Soper</name>
<name sortKey="Soper, Steven A" sort="Soper, Steven A" uniqKey="Soper S" first="Steven A." last="Soper">Steven A. Soper</name>
<name sortKey="Soper, Steven A" sort="Soper, Steven A" uniqKey="Soper S" first="Steven A." last="Soper">Steven A. Soper</name>
<name sortKey="Wheatley, Abigail" sort="Wheatley, Abigail" uniqKey="Wheatley A" first="Abigail" last="Wheatley">Abigail Wheatley</name>
<name sortKey="Witek, Malgorzata A" sort="Witek, Malgorzata A" uniqKey="Witek M" first="Małgorzata A." last="Witek">Małgorzata A. Witek</name>
</country>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Belgique/explor/OpenAccessBelV2/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001625 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 001625 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Wicri/Belgique |area= OpenAccessBelV2 |flux= Main |étape= Exploration |type= RBID |clé= ISTEX:F204A775A297E50260E05CE15E94036537BAC12F |texte= Purification and preconcentration of genomic DNA from whole cell lysates using photoactivated polycarbonate (PPC) microfluidic chips }}
This area was generated with Dilib version V0.6.25. |