Separation of oligonucleotides of identical size, but different base composition, by free zone capillary electrophoresis in strongly acidic, isoelectric buffers.
Identifieur interne : 002B05 ( Ncbi/Merge ); précédent : 002B04; suivant : 002B06Separation of oligonucleotides of identical size, but different base composition, by free zone capillary electrophoresis in strongly acidic, isoelectric buffers.
Auteurs : M. Perego [Italie] ; C. Gelfi ; A V Stoyanov ; P G RighettiSource :
- Electrophoresis [ 0173-0835 ] ; 1997.
Descripteurs français
- KwdFr :
- MESH :
- isolement et purification : Oligonucléotides.
- Acides, Composition en bases nucléiques, Concentration en ions d'hydrogène, Point isoélectrique, Substances tampon, Titrimétrie, Électrophorèse capillaire.
English descriptors
- KwdEn :
- MESH :
- chemical , isolation & purification : Oligonucleotides.
- chemical : Acids, Buffers.
- methods : Electrophoresis, Capillary.
- Base Composition, Hydrogen-Ion Concentration, Isoelectric Point, Titrimetry.
Abstract
A novel method for analyzing oligonucleotides of the same length, but bearing a single base substitution, is reported, based on free zone capillary electrophoresis (CZE) under rather acidic pH values. For this purpose, a set of four 18-mers of fairly random base composition has been synthesized, bearing, in nucleotide 9, the following bases: T, C, G or A. Theoretical predictions, based on titration curves of single free nucleotides, allowed us to predict that the simultaneous separation of a mixture of all four oligonucleotides could be possible in a pH 3-4 window. In fact, electrophoresis at pH 5.7 gave a single, asymmetric peak, whereas CZE at pH 4.8 could resolve three out of four species (the T9 and G9 oligonucleotides co-migrating into a single zone). A unique separation power could be obtained at pH 3.3 in a buffer comprising an amphoteric species (isoelectric iminodiacetic acid, IDA) and 7 M urea. Although IDA exhibited a pI of 2.23 (for a 100 mM solution), the addition of 7 M urea (necessary to denature the oligonucleotides) raised the apparent pH of the solution to 3.3.
DOI: 10.1002/elps.1150181532
PubMed: 9504830
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pubmed:9504830Le document en format XML
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Gelfi</name></author><author><name sortKey="Stoyanov, A V" sort="Stoyanov, A V" uniqKey="Stoyanov A" first="A V" last="Stoyanov">A V Stoyanov</name></author><author><name sortKey="Righetti, P G" sort="Righetti, P G" uniqKey="Righetti P" first="P G" last="Righetti">P G Righetti</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1997">1997</date><idno type="RBID">pubmed:9504830</idno><idno type="pmid">9504830</idno><idno type="doi">10.1002/elps.1150181532</idno><idno type="wicri:Area/PubMed/Corpus">002700</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002700</idno><idno type="wicri:Area/PubMed/Curation">002700</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002700</idno><idno type="wicri:Area/PubMed/Checkpoint">002570</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002570</idno><idno type="wicri:Area/Ncbi/Merge">002B05</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Separation of oligonucleotides of identical size, but different base composition, by free zone capillary electrophoresis in strongly acidic, isoelectric buffers.</title><author><name sortKey="Perego, M" sort="Perego, M" uniqKey="Perego M" first="M" last="Perego">M. Perego</name><affiliation wicri:level="1"><nlm:affiliation>Department of Agricultural and Industrial Biotechnologies, University of Verona, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Agricultural and Industrial Biotechnologies, University of Verona</wicri:regionArea><wicri:noRegion>University of Verona</wicri:noRegion></affiliation></author><author><name sortKey="Gelfi, C" sort="Gelfi, C" uniqKey="Gelfi C" first="C" last="Gelfi">C. Gelfi</name></author><author><name sortKey="Stoyanov, A V" sort="Stoyanov, A V" uniqKey="Stoyanov A" first="A V" last="Stoyanov">A V Stoyanov</name></author><author><name sortKey="Righetti, P G" sort="Righetti, P G" uniqKey="Righetti P" first="P G" last="Righetti">P G Righetti</name></author></analytic><series><title level="j">Electrophoresis</title><idno type="ISSN">0173-0835</idno><imprint><date when="1997" type="published">1997</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Acids</term><term>Base Composition</term><term>Buffers</term><term>Electrophoresis, Capillary (methods)</term><term>Hydrogen-Ion Concentration</term><term>Isoelectric Point</term><term>Oligonucleotides (isolation & purification)</term><term>Titrimetry</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Acides</term><term>Composition en bases nucléiques</term><term>Concentration en ions d'hydrogène</term><term>Oligonucléotides (isolement et purification)</term><term>Point isoélectrique</term><term>Substances tampon</term><term>Titrimétrie</term><term>Électrophorèse capillaire ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Oligonucleotides</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Acids</term><term>Buffers</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Oligonucléotides</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Electrophoresis, Capillary</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Composition</term><term>Hydrogen-Ion Concentration</term><term>Isoelectric Point</term><term>Titrimetry</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Acides</term><term>Composition en bases nucléiques</term><term>Concentration en ions d'hydrogène</term><term>Point isoélectrique</term><term>Substances tampon</term><term>Titrimétrie</term><term>Électrophorèse capillaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A novel method for analyzing oligonucleotides of the same length, but bearing a single base substitution, is reported, based on free zone capillary electrophoresis (CZE) under rather acidic pH values. For this purpose, a set of four 18-mers of fairly random base composition has been synthesized, bearing, in nucleotide 9, the following bases: T, C, G or A. Theoretical predictions, based on titration curves of single free nucleotides, allowed us to predict that the simultaneous separation of a mixture of all four oligonucleotides could be possible in a pH 3-4 window. In fact, electrophoresis at pH 5.7 gave a single, asymmetric peak, whereas CZE at pH 4.8 could resolve three out of four species (the T9 and G9 oligonucleotides co-migrating into a single zone). A unique separation power could be obtained at pH 3.3 in a buffer comprising an amphoteric species (isoelectric iminodiacetic acid, IDA) and 7 M urea. Although IDA exhibited a pI of 2.23 (for a 100 mM solution), the addition of 7 M urea (necessary to denature the oligonucleotides) raised the apparent pH of the solution to 3.3.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">9504830</PMID><DateCompleted><Year>1998</Year><Month>05</Month><Day>01</Day></DateCompleted><DateRevised><Year>2006</Year><Month>11</Month><Day>15</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0173-0835</ISSN><JournalIssue CitedMedium="Print"><Volume>18</Volume><Issue>15</Issue><PubDate><Year>1997</Year><Month>Dec</Month></PubDate></JournalIssue><Title>Electrophoresis</Title><ISOAbbreviation>Electrophoresis</ISOAbbreviation></Journal><ArticleTitle>Separation of oligonucleotides of identical size, but different base composition, by free zone capillary electrophoresis in strongly acidic, isoelectric buffers.</ArticleTitle><Pagination><MedlinePgn>2915-20</MedlinePgn></Pagination><Abstract><AbstractText>A novel method for analyzing oligonucleotides of the same length, but bearing a single base substitution, is reported, based on free zone capillary electrophoresis (CZE) under rather acidic pH values. For this purpose, a set of four 18-mers of fairly random base composition has been synthesized, bearing, in nucleotide 9, the following bases: T, C, G or A. Theoretical predictions, based on titration curves of single free nucleotides, allowed us to predict that the simultaneous separation of a mixture of all four oligonucleotides could be possible in a pH 3-4 window. In fact, electrophoresis at pH 5.7 gave a single, asymmetric peak, whereas CZE at pH 4.8 could resolve three out of four species (the T9 and G9 oligonucleotides co-migrating into a single zone). A unique separation power could be obtained at pH 3.3 in a buffer comprising an amphoteric species (isoelectric iminodiacetic acid, IDA) and 7 M urea. Although IDA exhibited a pI of 2.23 (for a 100 mM solution), the addition of 7 M urea (necessary to denature the oligonucleotides) raised the apparent pH of the solution to 3.3.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Perego</LastName><ForeName>M</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Agricultural and Industrial Biotechnologies, University of Verona, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Gelfi</LastName><ForeName>C</ForeName><Initials>C</Initials></Author><Author ValidYN="Y"><LastName>Stoyanov</LastName><ForeName>A V</ForeName><Initials>AV</Initials></Author><Author ValidYN="Y"><LastName>Righetti</LastName><ForeName>P G</ForeName><Initials>PG</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D003160">Comparative Study</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Electrophoresis</MedlineTA><NlmUniqueID>8204476</NlmUniqueID><ISSNLinking>0173-0835</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000143">Acids</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D002021">Buffers</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009841">Oligonucleotides</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000143" MajorTopicYN="N">Acids</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001482" MajorTopicYN="Y">Base Composition</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002021" MajorTopicYN="N">Buffers</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019075" MajorTopicYN="N">Electrophoresis, Capillary</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006863" MajorTopicYN="N">Hydrogen-Ion Concentration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007526" MajorTopicYN="N">Isoelectric Point</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009841" MajorTopicYN="N">Oligonucleotides</DescriptorName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D017186" MajorTopicYN="N">Titrimetry</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1998</Year><Month>3</Month><Day>21</Day></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>1998</Year><Month>3</Month><Day>21</Day><Hour>0</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>1998</Year><Month>3</Month><Day>21</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">9504830</ArticleId><ArticleId IdType="doi">10.1002/elps.1150181532</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Italie</li></country></list><tree><noCountry><name sortKey="Gelfi, C" sort="Gelfi, C" uniqKey="Gelfi C" first="C" last="Gelfi">C. Gelfi</name><name sortKey="Righetti, P G" sort="Righetti, P G" uniqKey="Righetti P" first="P G" last="Righetti">P G Righetti</name><name sortKey="Stoyanov, A V" sort="Stoyanov, A V" uniqKey="Stoyanov A" first="A V" last="Stoyanov">A V Stoyanov</name></noCountry><country name="Italie"><noRegion><name sortKey="Perego, M" sort="Perego, M" uniqKey="Perego M" first="M" last="Perego">M. Perego</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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