Probe generation directly from small numbers of cells for DNA microarray studies.
Identifieur interne : 002468 ( PubMed/Curation ); précédent : 002467; suivant : 002469Probe generation directly from small numbers of cells for DNA microarray studies.
Auteurs : C C Xiang [États-Unis] ; M. Chen ; O A Kozhich ; Q N Phan ; J M Inman ; Y. Chen ; M J BrownsteinSource :
- BioTechniques [ 0736-6205 ] ; 2003.
Descripteurs français
- KwdFr :
- ARN (analyse), ARN (génétique), Analyse de profil d'expression de gènes (), Animaux, Cellules 3T3, Contrôle de qualité, Données de séquences moléculaires, Microchimie (), Microchimie (instrumentation), Sondes d'ADN (biosynthèse), Sondes d'ADN (synthèse chimique), Souris, Séquence nucléotidique, Séquençage par oligonucléotides en batterie (), Séquençage par oligonucléotides en batterie (instrumentation), Techniques d'amplification d'acides nucléiques ().
- MESH :
- analyse : ARN.
- biosynthèse : Sondes d'ADN.
- génétique : ARN.
- synthèse chimique : Microchimie, Sondes d'ADN, Séquençage par oligonucléotides en batterie.
- Analyse de profil d'expression de gènes, Animaux, Cellules 3T3, Contrôle de qualité, Données de séquences moléculaires, Microchimie, Souris, Séquence nucléotidique, Séquençage par oligonucléotides en batterie, Techniques d'amplification d'acides nucléiques.
English descriptors
- KwdEn :
- 3T3 Cells, Animals, Base Sequence, DNA Probes (biosynthesis), DNA Probes (chemical synthesis), Gene Expression Profiling (methods), Mice, Microchemistry (instrumentation), Microchemistry (methods), Molecular Sequence Data, Nucleic Acid Amplification Techniques (methods), Oligonucleotide Array Sequence Analysis (instrumentation), Oligonucleotide Array Sequence Analysis (methods), Quality Control, RNA (analysis), RNA (genetics).
- MESH :
- chemical , analysis : RNA.
- chemical , biosynthesis : DNA Probes.
- chemical , chemical synthesis : DNA Probes.
- chemical , genetics : RNA.
- instrumentation : Microchemistry, Oligonucleotide Array Sequence Analysis.
- methods : Gene Expression Profiling, Microchemistry, Nucleic Acid Amplification Techniques, Oligonucleotide Array Sequence Analysis.
- 3T3 Cells, Animals, Base Sequence, Mice, Molecular Sequence Data, Quality Control.
Abstract
Recently, we described a technique that allows us to prepare probes for expression profiling from 0.5-1 microgram RNA without template or signal amplification. However, we were unable to use this method to study cells harvested by needle biopsy, cell sorting, or laser capture microdissection. Here we give a new protocol for amplifying RNA with multiple reaction cycles and preparing fluorescent probes from approximately 10 cells. We use random 9-mers with a T3 RNA polymerase recognition sequence on the 5' end for every round of cDNA synthesis except the first. The latter is primed with oligo(dT) with a T7 RNA polymerase recognition sequence on the 5' end. Results were highly reproducible and reliable, and the products generated using our method seemed comparable to those produced using the RiboAmp RNA kit when both were used to do two cycles of amplification. To test our method's utility, we lysed cells directly into reverse transcription buffer containing RNase inhibitor and performed three rounds of RNA amplification. The expression profiles of mouse C2 and NIH 3T3 cells obtained with 11,232-element arrays using amplified RNAs were similar to those seen when probes were prepared from unamplified templates.
DOI: 10.2144/03342mt03
PubMed: 12613261
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: Pour aller vers cette notice dans l'étape Curation :002468
Links to Exploration step
pubmed:12613261Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Probe generation directly from small numbers of cells for DNA microarray studies.</title><author><name sortKey="Xiang, C C" sort="Xiang, C C" uniqKey="Xiang C" first="C C" last="Xiang">C C Xiang</name><affiliation wicri:level="1"><nlm:affiliation>NIMH/NIH, Bethesda, MD, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>NIMH/NIH, Bethesda, MD</wicri:regionArea></affiliation></author><author><name sortKey="Chen, M" sort="Chen, M" uniqKey="Chen M" first="M" last="Chen">M. Chen</name></author><author><name sortKey="Kozhich, O A" sort="Kozhich, O A" uniqKey="Kozhich O" first="O A" last="Kozhich">O A Kozhich</name></author><author><name sortKey="Phan, Q N" sort="Phan, Q N" uniqKey="Phan Q" first="Q N" last="Phan">Q N Phan</name></author><author><name sortKey="Inman, J M" sort="Inman, J M" uniqKey="Inman J" first="J M" last="Inman">J M Inman</name></author><author><name sortKey="Chen, Y" sort="Chen, Y" uniqKey="Chen Y" first="Y" last="Chen">Y. Chen</name></author><author><name sortKey="Brownstein, M J" sort="Brownstein, M J" uniqKey="Brownstein M" first="M J" last="Brownstein">M J Brownstein</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2003">2003</date><idno type="RBID">pubmed:12613261</idno><idno type="pmid">12613261</idno><idno type="doi">10.2144/03342mt03</idno><idno type="wicri:Area/PubMed/Corpus">002468</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002468</idno><idno type="wicri:Area/PubMed/Curation">002468</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002468</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Probe generation directly from small numbers of cells for DNA microarray studies.</title><author><name sortKey="Xiang, C C" sort="Xiang, C C" uniqKey="Xiang C" first="C C" last="Xiang">C C Xiang</name><affiliation wicri:level="1"><nlm:affiliation>NIMH/NIH, Bethesda, MD, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>NIMH/NIH, Bethesda, MD</wicri:regionArea></affiliation></author><author><name sortKey="Chen, M" sort="Chen, M" uniqKey="Chen M" first="M" last="Chen">M. Chen</name></author><author><name sortKey="Kozhich, O A" sort="Kozhich, O A" uniqKey="Kozhich O" first="O A" last="Kozhich">O A Kozhich</name></author><author><name sortKey="Phan, Q N" sort="Phan, Q N" uniqKey="Phan Q" first="Q N" last="Phan">Q N Phan</name></author><author><name sortKey="Inman, J M" sort="Inman, J M" uniqKey="Inman J" first="J M" last="Inman">J M Inman</name></author><author><name sortKey="Chen, Y" sort="Chen, Y" uniqKey="Chen Y" first="Y" last="Chen">Y. Chen</name></author><author><name sortKey="Brownstein, M J" sort="Brownstein, M J" uniqKey="Brownstein M" first="M J" last="Brownstein">M J Brownstein</name></author></analytic><series><title level="j">BioTechniques</title><idno type="ISSN">0736-6205</idno><imprint><date when="2003" type="published">2003</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>3T3 Cells</term><term>Animals</term><term>Base Sequence</term><term>DNA Probes (biosynthesis)</term><term>DNA Probes (chemical synthesis)</term><term>Gene Expression Profiling (methods)</term><term>Mice</term><term>Microchemistry (instrumentation)</term><term>Microchemistry (methods)</term><term>Molecular Sequence Data</term><term>Nucleic Acid Amplification Techniques (methods)</term><term>Oligonucleotide Array Sequence Analysis (instrumentation)</term><term>Oligonucleotide Array Sequence Analysis (methods)</term><term>Quality Control</term><term>RNA (analysis)</term><term>RNA (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN (analyse)</term><term>ARN (génétique)</term><term>Analyse de profil d'expression de gènes ()</term><term>Animaux</term><term>Cellules 3T3</term><term>Contrôle de qualité</term><term>Données de séquences moléculaires</term><term>Microchimie ()</term><term>Microchimie (instrumentation)</term><term>Sondes d'ADN (biosynthèse)</term><term>Sondes d'ADN (synthèse chimique)</term><term>Souris</term><term>Séquence nucléotidique</term><term>Séquençage par oligonucléotides en batterie ()</term><term>Séquençage par oligonucléotides en batterie (instrumentation)</term><term>Techniques d'amplification d'acides nucléiques ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>RNA</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>DNA Probes</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemical synthesis" xml:lang="en"><term>DNA Probes</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ARN</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Sondes d'ADN</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN</term></keywords><keywords scheme="MESH" qualifier="instrumentation" xml:lang="en"><term>Microchemistry</term><term>Oligonucleotide Array Sequence Analysis</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Gene Expression Profiling</term><term>Microchemistry</term><term>Nucleic Acid Amplification Techniques</term><term>Oligonucleotide Array Sequence Analysis</term></keywords><keywords scheme="MESH" qualifier="synthèse chimique" xml:lang="fr"><term>Microchimie</term><term>Sondes d'ADN</term><term>Séquençage par oligonucléotides en batterie</term></keywords><keywords scheme="MESH" xml:lang="en"><term>3T3 Cells</term><term>Animals</term><term>Base Sequence</term><term>Mice</term><term>Molecular Sequence Data</term><term>Quality Control</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse de profil d'expression de gènes</term><term>Animaux</term><term>Cellules 3T3</term><term>Contrôle de qualité</term><term>Données de séquences moléculaires</term><term>Microchimie</term><term>Souris</term><term>Séquence nucléotidique</term><term>Séquençage par oligonucléotides en batterie</term><term>Techniques d'amplification d'acides nucléiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Recently, we described a technique that allows us to prepare probes for expression profiling from 0.5-1 microgram RNA without template or signal amplification. However, we were unable to use this method to study cells harvested by needle biopsy, cell sorting, or laser capture microdissection. Here we give a new protocol for amplifying RNA with multiple reaction cycles and preparing fluorescent probes from approximately 10 cells. We use random 9-mers with a T3 RNA polymerase recognition sequence on the 5' end for every round of cDNA synthesis except the first. The latter is primed with oligo(dT) with a T7 RNA polymerase recognition sequence on the 5' end. Results were highly reproducible and reliable, and the products generated using our method seemed comparable to those produced using the RiboAmp RNA kit when both were used to do two cycles of amplification. To test our method's utility, we lysed cells directly into reverse transcription buffer containing RNase inhibitor and performed three rounds of RNA amplification. The expression profiles of mouse C2 and NIH 3T3 cells obtained with 11,232-element arrays using amplified RNAs were similar to those seen when probes were prepared from unamplified templates.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">12613261</PMID><DateCompleted><Year>2003</Year><Month>08</Month><Day>15</Day></DateCompleted><DateRevised><Year>2019</Year><Month>12</Month><Day>10</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0736-6205</ISSN><JournalIssue CitedMedium="Print"><Volume>34</Volume><Issue>2</Issue><PubDate><Year>2003</Year><Month>Feb</Month></PubDate></JournalIssue><Title>BioTechniques</Title><ISOAbbreviation>BioTechniques</ISOAbbreviation></Journal><ArticleTitle>Probe generation directly from small numbers of cells for DNA microarray studies.</ArticleTitle><Pagination><MedlinePgn>386-8, 390, 392-3</MedlinePgn></Pagination><Abstract><AbstractText>Recently, we described a technique that allows us to prepare probes for expression profiling from 0.5-1 microgram RNA without template or signal amplification. However, we were unable to use this method to study cells harvested by needle biopsy, cell sorting, or laser capture microdissection. Here we give a new protocol for amplifying RNA with multiple reaction cycles and preparing fluorescent probes from approximately 10 cells. We use random 9-mers with a T3 RNA polymerase recognition sequence on the 5' end for every round of cDNA synthesis except the first. The latter is primed with oligo(dT) with a T7 RNA polymerase recognition sequence on the 5' end. Results were highly reproducible and reliable, and the products generated using our method seemed comparable to those produced using the RiboAmp RNA kit when both were used to do two cycles of amplification. To test our method's utility, we lysed cells directly into reverse transcription buffer containing RNase inhibitor and performed three rounds of RNA amplification. The expression profiles of mouse C2 and NIH 3T3 cells obtained with 11,232-element arrays using amplified RNAs were similar to those seen when probes were prepared from unamplified templates.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Xiang</LastName><ForeName>C C</ForeName><Initials>CC</Initials><AffiliationInfo><Affiliation>NIMH/NIH, Bethesda, MD, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>M</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Kozhich</LastName><ForeName>O A</ForeName><Initials>OA</Initials></Author><Author ValidYN="Y"><LastName>Phan</LastName><ForeName>Q N</ForeName><Initials>QN</Initials></Author><Author ValidYN="Y"><LastName>Inman</LastName><ForeName>J M</ForeName><Initials>JM</Initials></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>Y</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Brownstein</LastName><ForeName>M J</ForeName><Initials>MJ</Initials></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>N01-CO-12400.</GrantID><Acronym>CO</Acronym><Agency>NCI NIH HHS</Agency><Country>United States</Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D023362">Evaluation Study</PublicationType><PublicationType UI="D013487">Research Support, U.S. Gov't, P.H.S.</PublicationType><PublicationType UI="D016427">Technical Report</PublicationType><PublicationType UI="D023361">Validation Study</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Biotechniques</MedlineTA><NlmUniqueID>8306785</NlmUniqueID><ISSNLinking>0736-6205</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D015342">DNA Probes</NameOfSubstance></Chemical><Chemical><RegistryNumber>63231-63-0</RegistryNumber><NameOfSubstance UI="D012313">RNA</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D016475" MajorTopicYN="N">3T3 Cells</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015342" MajorTopicYN="N">DNA Probes</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000138" MajorTopicYN="Y">chemical synthesis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020869" MajorTopicYN="N">Gene Expression Profiling</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D051379" MajorTopicYN="N">Mice</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008832" MajorTopicYN="N">Microchemistry</DescriptorName><QualifierName UI="Q000295" MajorTopicYN="N">instrumentation</QualifierName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D021141" MajorTopicYN="N">Nucleic Acid Amplification Techniques</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020411" MajorTopicYN="N">Oligonucleotide Array Sequence Analysis</DescriptorName><QualifierName UI="Q000295" MajorTopicYN="N">instrumentation</QualifierName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011786" MajorTopicYN="N">Quality Control</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012313" MajorTopicYN="N">RNA</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2003</Year><Month>3</Month><Day>5</Day><Hour>4</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2003</Year><Month>8</Month><Day>16</Day><Hour>5</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2003</Year><Month>3</Month><Day>5</Day><Hour>4</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">12613261</ArticleId><ArticleId IdType="doi">10.2144/03342mt03</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/PubMed/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002468 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/PubMed/Curation/biblio.hfd -nk 002468 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= PubMed
|étape= Curation
|type= RBID
|clé= pubmed:12613261
|texte= Probe generation directly from small numbers of cells for DNA microarray studies.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/PubMed/Curation/RBID.i -Sk "pubmed:12613261" \
| HfdSelect -Kh $EXPLOR_AREA/Data/PubMed/Curation/biblio.hfd \
| NlmPubMed2Wicri -a MersV1
| This area was generated with Dilib version V0.6.33. |  |