Analysis and purification of peptide nucleic acids by denaturing PAGE.
Identifieur interne : 002077 ( PubMed/Checkpoint ); précédent : 002076; suivant : 002078Analysis and purification of peptide nucleic acids by denaturing PAGE.
Auteurs : David W. Dodd [Canada] ; Robert H E. HudsonSource :
- Electrophoresis [ 0173-0835 ] ; 2007.
Descripteurs français
- KwdFr :
- ADN (), ADN (isolement et purification), Acides nucléiques peptidiques (), Acides nucléiques peptidiques (isolement et purification), Appariement de bases, Concentration en ions d'hydrogène, Dénaturation d'acide nucléique, Dénaturation des protéines, Masse moléculaire, Oligonucléotides (), Oligonucléotides (isolement et purification), Sensibilité et spécificité, Spectrophotométrie UV, Séquence nucléotidique, Urée (), Électrophorèse sur gel de polyacrylamide ().
- MESH :
- isolement et purification : ADN, Acides nucléiques peptidiques, Oligonucléotides.
- ADN, Acides nucléiques peptidiques, Appariement de bases, Concentration en ions d'hydrogène, Dénaturation d'acide nucléique, Dénaturation des protéines, Masse moléculaire, Oligonucléotides, Sensibilité et spécificité, Spectrophotométrie UV, Séquence nucléotidique, Urée, Électrophorèse sur gel de polyacrylamide.
English descriptors
- KwdEn :
- Base Pairing, Base Sequence, DNA (chemistry), DNA (isolation & purification), Electrophoresis, Polyacrylamide Gel (methods), Hydrogen-Ion Concentration, Molecular Weight, Nucleic Acid Denaturation, Oligonucleotides (chemistry), Oligonucleotides (isolation & purification), Peptide Nucleic Acids (chemistry), Peptide Nucleic Acids (isolation & purification), Protein Denaturation, Sensitivity and Specificity, Spectrophotometry, Ultraviolet, Urea (chemistry).
- MESH :
- chemical , chemistry : DNA, Oligonucleotides, Peptide Nucleic Acids, Urea.
- chemical , isolation & purification : DNA, Oligonucleotides, Peptide Nucleic Acids.
- methods : Electrophoresis, Polyacrylamide Gel.
- Base Pairing, Base Sequence, Hydrogen-Ion Concentration, Molecular Weight, Nucleic Acid Denaturation, Protein Denaturation, Sensitivity and Specificity, Spectrophotometry, Ultraviolet.
Abstract
A flexible and convenient protocol for the analysis and purification of peptide nucleic acid (PNA) oligomers and PNA-peptide chimeras by denaturing PAGE is described. Vertical slab gel electrophoresis, 26% in polyacrylamide and 8 M urea at pH 3, was suitable for analysis of oligomers ranging in size from tetramers (4-mers) to tetradodecamers (24-mers). Single-base resolution of oligomers was achieved and separations are generally superior to those given by standard RP-HPLC techniques. The separation of a related series of PNA oligomers showed the distance migrated was linearly dependent on the logarithm of the molecular weight. The migration of oligomers through the gel is dependent on the number of basic functional groups present, such as amino groups, and the A and C content of the oligomer. PNAs are amenable to detection by UV-shadowing technique illuminated at 260 nm or Coomassie blue staining, both with similar, sub-microgram per band detection limits.
DOI: 10.1002/elps.200700192
PubMed: 17922504
Affiliations:
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Vertical slab gel electrophoresis, 26% in polyacrylamide and 8 M urea at pH 3, was suitable for analysis of oligomers ranging in size from tetramers (4-mers) to tetradodecamers (24-mers). Single-base resolution of oligomers was achieved and separations are generally superior to those given by standard RP-HPLC techniques. The separation of a related series of PNA oligomers showed the distance migrated was linearly dependent on the logarithm of the molecular weight. The migration of oligomers through the gel is dependent on the number of basic functional groups present, such as amino groups, and the A and C content of the oligomer. PNAs are amenable to detection by UV-shadowing technique illuminated at 260 nm or Coomassie blue staining, both with similar, sub-microgram per band detection limits.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">17922504</PMID><DateCompleted><Year>2008</Year><Month>02</Month><Day>26</Day></DateCompleted><DateRevised><Year>2013</Year><Month>11</Month><Day>21</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0173-0835</ISSN><JournalIssue CitedMedium="Print"><Volume>28</Volume><Issue>21</Issue><PubDate><Year>2007</Year><Month>Nov</Month></PubDate></JournalIssue><Title>Electrophoresis</Title><ISOAbbreviation>Electrophoresis</ISOAbbreviation></Journal><ArticleTitle>Analysis and purification of peptide nucleic acids by denaturing PAGE.</ArticleTitle><Pagination><MedlinePgn>3884-9</MedlinePgn></Pagination><Abstract><AbstractText>A flexible and convenient protocol for the analysis and purification of peptide nucleic acid (PNA) oligomers and PNA-peptide chimeras by denaturing PAGE is described. Vertical slab gel electrophoresis, 26% in polyacrylamide and 8 M urea at pH 3, was suitable for analysis of oligomers ranging in size from tetramers (4-mers) to tetradodecamers (24-mers). Single-base resolution of oligomers was achieved and separations are generally superior to those given by standard RP-HPLC techniques. The separation of a related series of PNA oligomers showed the distance migrated was linearly dependent on the logarithm of the molecular weight. The migration of oligomers through the gel is dependent on the number of basic functional groups present, such as amino groups, and the A and C content of the oligomer. PNAs are amenable to detection by UV-shadowing technique illuminated at 260 nm or Coomassie blue staining, both with similar, sub-microgram per band detection limits.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Dodd</LastName><ForeName>David W</ForeName><Initials>DW</Initials><AffiliationInfo><Affiliation>Department of Chemistry, The University of Western Ontario, London, ON, Canada.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hudson</LastName><ForeName>Robert H E</ForeName><Initials>RH</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><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="D009841">Oligonucleotides</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D020135">Peptide Nucleic Acids</NameOfSubstance></Chemical><Chemical><RegistryNumber>8W8T17847W</RegistryNumber><NameOfSubstance UI="D014508">Urea</NameOfSubstance></Chemical><Chemical><RegistryNumber>9007-49-2</RegistryNumber><NameOfSubstance UI="D004247">DNA</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D020029" MajorTopicYN="N">Base Pairing</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004247" MajorTopicYN="N">DNA</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004591" MajorTopicYN="N">Electrophoresis, Polyacrylamide Gel</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006863" MajorTopicYN="N">Hydrogen-Ion Concentration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008970" MajorTopicYN="N">Molecular Weight</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009691" MajorTopicYN="Y">Nucleic Acid Denaturation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009841" MajorTopicYN="N">Oligonucleotides</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020135" MajorTopicYN="N">Peptide Nucleic Acids</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011489" MajorTopicYN="N">Protein Denaturation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012680" MajorTopicYN="N">Sensitivity and Specificity</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013056" MajorTopicYN="N">Spectrophotometry, Ultraviolet</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014508" MajorTopicYN="N">Urea</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2007</Year><Month>10</Month><Day>9</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2008</Year><Month>2</Month><Day>27</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2007</Year><Month>10</Month><Day>9</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">17922504</ArticleId><ArticleId IdType="doi">10.1002/elps.200700192</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Canada</li></country></list><tree><noCountry><name sortKey="Hudson, Robert H E" sort="Hudson, Robert H E" uniqKey="Hudson R" first="Robert H E" last="Hudson">Robert H E. Hudson</name></noCountry><country name="Canada"><noRegion><name sortKey="Dodd, David W" sort="Dodd, David W" uniqKey="Dodd D" first="David W" last="Dodd">David W. Dodd</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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