A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants.
Identifieur interne : 003842 ( Main/Curation ); précédent : 003841; suivant : 003843A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants.
Auteurs : Giorgio Gambino [Italie] ; Irene Perrone ; Ivana GribaudoSource :
- Phytochemical analysis : PCA [ 1099-1565 ]
Descripteurs français
- KwdFr :
- ARN des plantes (isolement et purification), Arbres (composition chimique), Arbres (génétique), Bromure de cétrimonium (MeSH), Composés de cétrimonium (composition chimique), Feuilles de plante (composition chimique), Feuilles de plante (génétique), RT-PCR (MeSH), Techniques de chimie analytique (méthodes), Techniques de chimie analytique (économie), Vitis (composition chimique), Vitis (génétique), Électrophorèse sur gel d'agar (MeSH).
- MESH :
- composition chimique : Arbres, Composés de cétrimonium, Feuilles de plante, Vitis.
- génétique : Arbres, Feuilles de plante, Vitis.
- isolement et purification : ARN des plantes.
- méthodes : Techniques de chimie analytique.
- économie : Techniques de chimie analytique.
- Bromure de cétrimonium, RT-PCR, Électrophorèse sur gel d'agar.
English descriptors
- KwdEn :
- Cetrimonium (MeSH), Cetrimonium Compounds (chemistry), Chemistry Techniques, Analytical (economics), Chemistry Techniques, Analytical (methods), Electrophoresis, Agar Gel (MeSH), Plant Leaves (chemistry), Plant Leaves (genetics), RNA, Plant (isolation & purification), Reverse Transcriptase Polymerase Chain Reaction (MeSH), Trees (chemistry), Trees (genetics), Vitis (chemistry), Vitis (genetics).
- MESH :
- chemical , chemistry : Cetrimonium Compounds.
- chemical , isolation & purification : RNA, Plant.
- chemical : Cetrimonium.
- chemistry : Plant Leaves, Trees, Vitis.
- economics : Chemistry Techniques, Analytical.
- genetics : Plant Leaves, Trees, Vitis.
- methods : Chemistry Techniques, Analytical.
- Electrophoresis, Agar Gel, Reverse Transcriptase Polymerase Chain Reaction.
Abstract
INTRODUCTION
RNA quality and integrity are critical for many studies in plant molecular biology. High-quality RNA extraction from grapevine and other woody plants is problematic due to the presence of polysaccharides, polyphenolics and other compounds that bind or co-precipitate with the RNA.
OBJECTIVE
To develop an optimised cetyltrimethylammonium bromide (CTAB)-based protocol, to reduce the time and cost of extraction without reducing quality and yield of RNA extracted from polysaccharide-rich tissues of several plants.
METHODOLOGY
Several changes were introduced to the original CTAB protocol. All centrifugation steps were carried out at 4 degrees C, the sample weight was decreased and the concentrations of PVP-40 and LiCl were increased reducing incubation time prior to RNA precipitation. This rapid CTAB protocol was compared with six different RNA extraction methods from three grapevine tissues, namely, in vitro plantlets, and leaves and mature canes from actively growing field vines.
RESULTS
The rapid CTAB method gave high-quality RNA in only 3 h at low cost with efficiency equal to or higher than that obtained with other time-consuming and expensive protocols. The procedure was applied to RNA extraction from other grapevine tissues and other woody species including olive, lemon, poplar, chestnut, apple, pear, peach, cherry, apricot, plum and kiwi fruit. RNA of high quality could be isolated from all tissues and from all species.
CONCLUSION
The study has shown that the improvement of a CTAB-based protocol allows the rapid isolation of high-quality RNA from grapevine and many woody species.
DOI: 10.1002/pca.1078
PubMed: 18618437
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Via L. da Vinci 44, I-10095 Grugliasco-TO</wicri:regionArea></affiliation></author><author><name sortKey="Perrone, Irene" sort="Perrone, Irene" uniqKey="Perrone I" first="Irene" last="Perrone">Irene Perrone</name></author><author><name sortKey="Gribaudo, Ivana" sort="Gribaudo, Ivana" uniqKey="Gribaudo I" first="Ivana" last="Gribaudo">Ivana Gribaudo</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2008">2008 Nov-Dec</date><idno type="RBID">pubmed:18618437</idno><idno type="pmid">18618437</idno><idno type="doi">10.1002/pca.1078</idno><idno type="wicri:Area/Main/Corpus">003842</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">003842</idno><idno type="wicri:Area/Main/Curation">003842</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">003842</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants.</title><author><name sortKey="Gambino, Giorgio" sort="Gambino, Giorgio" uniqKey="Gambino G" first="Giorgio" last="Gambino">Giorgio Gambino</name><affiliation wicri:level="1"><nlm:affiliation>Plant Virology Institute CNR, Grugliasco Unit. Via L. da Vinci 44, I-10095 Grugliasco-TO, Italy. g.gambino@ivv.cnr.it</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Plant Virology Institute CNR, Grugliasco Unit. 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High-quality RNA extraction from grapevine and other woody plants is problematic due to the presence of polysaccharides, polyphenolics and other compounds that bind or co-precipitate with the RNA.</p></div><div type="abstract" xml:lang="en"><p><b>OBJECTIVE</b></p><p>To develop an optimised cetyltrimethylammonium bromide (CTAB)-based protocol, to reduce the time and cost of extraction without reducing quality and yield of RNA extracted from polysaccharide-rich tissues of several plants.</p></div><div type="abstract" xml:lang="en"><p><b>METHODOLOGY</b></p><p>Several changes were introduced to the original CTAB protocol. All centrifugation steps were carried out at 4 degrees C, the sample weight was decreased and the concentrations of PVP-40 and LiCl were increased reducing incubation time prior to RNA precipitation. This rapid CTAB protocol was compared with six different RNA extraction methods from three grapevine tissues, namely, in vitro plantlets, and leaves and mature canes from actively growing field vines.</p></div><div type="abstract" xml:lang="en"><p><b>RESULTS</b></p><p>The rapid CTAB method gave high-quality RNA in only 3 h at low cost with efficiency equal to or higher than that obtained with other time-consuming and expensive protocols. The procedure was applied to RNA extraction from other grapevine tissues and other woody species including olive, lemon, poplar, chestnut, apple, pear, peach, cherry, apricot, plum and kiwi fruit. RNA of high quality could be isolated from all tissues and from all species.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSION</b></p><p>The study has shown that the improvement of a CTAB-based protocol allows the rapid isolation of high-quality RNA from grapevine and many woody species.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">18618437</PMID><DateCompleted><Year>2009</Year><Month>01</Month><Day>09</Day></DateCompleted><DateRevised><Year>2019</Year><Month>12</Month><Day>10</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1099-1565</ISSN><JournalIssue CitedMedium="Internet"><Volume>19</Volume><Issue>6</Issue><PubDate><MedlineDate>2008 Nov-Dec</MedlineDate></PubDate></JournalIssue><Title>Phytochemical analysis : PCA</Title><ISOAbbreviation>Phytochem Anal</ISOAbbreviation></Journal><ArticleTitle>A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants.</ArticleTitle><Pagination><MedlinePgn>520-5</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1002/pca.1078</ELocationID><Abstract><AbstractText Label="INTRODUCTION" NlmCategory="BACKGROUND">RNA quality and integrity are critical for many studies in plant molecular biology. High-quality RNA extraction from grapevine and other woody plants is problematic due to the presence of polysaccharides, polyphenolics and other compounds that bind or co-precipitate with the RNA.</AbstractText><AbstractText Label="OBJECTIVE" NlmCategory="OBJECTIVE">To develop an optimised cetyltrimethylammonium bromide (CTAB)-based protocol, to reduce the time and cost of extraction without reducing quality and yield of RNA extracted from polysaccharide-rich tissues of several plants.</AbstractText><AbstractText Label="METHODOLOGY" NlmCategory="METHODS">Several changes were introduced to the original CTAB protocol. All centrifugation steps were carried out at 4 degrees C, the sample weight was decreased and the concentrations of PVP-40 and LiCl were increased reducing incubation time prior to RNA precipitation. This rapid CTAB protocol was compared with six different RNA extraction methods from three grapevine tissues, namely, in vitro plantlets, and leaves and mature canes from actively growing field vines.</AbstractText><AbstractText Label="RESULTS" NlmCategory="RESULTS">The rapid CTAB method gave high-quality RNA in only 3 h at low cost with efficiency equal to or higher than that obtained with other time-consuming and expensive protocols. The procedure was applied to RNA extraction from other grapevine tissues and other woody species including olive, lemon, poplar, chestnut, apple, pear, peach, cherry, apricot, plum and kiwi fruit. RNA of high quality could be isolated from all tissues and from all species.</AbstractText><AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">The study has shown that the improvement of a CTAB-based protocol allows the rapid isolation of high-quality RNA from grapevine and many woody species.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Gambino</LastName><ForeName>Giorgio</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>Plant Virology Institute CNR, Grugliasco Unit. Via L. da Vinci 44, I-10095 Grugliasco-TO, Italy. g.gambino@ivv.cnr.it</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Perrone</LastName><ForeName>Irene</ForeName><Initials>I</Initials></Author><Author ValidYN="Y"><LastName>Gribaudo</LastName><ForeName>Ivana</ForeName><Initials>I</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D003160">Comparative Study</PublicationType><PublicationType UI="D023362">Evaluation Study</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Phytochem Anal</MedlineTA><NlmUniqueID>9200492</NlmUniqueID><ISSNLinking>0958-0344</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D002593">Cetrimonium Compounds</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018749">RNA, Plant</NameOfSubstance></Chemical><Chemical><RegistryNumber>Z7FF1XKL7A</RegistryNumber><NameOfSubstance UI="D000077286">Cetrimonium</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000077286" MajorTopicYN="N">Cetrimonium</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002593" MajorTopicYN="N">Cetrimonium Compounds</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D002623" MajorTopicYN="N">Chemistry Techniques, Analytical</DescriptorName><QualifierName UI="Q000191" MajorTopicYN="N">economics</QualifierName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004587" MajorTopicYN="N">Electrophoresis, Agar Gel</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018515" MajorTopicYN="N">Plant Leaves</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018749" MajorTopicYN="N">RNA, Plant</DescriptorName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020133" MajorTopicYN="N">Reverse Transcriptase Polymerase Chain Reaction</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014197" MajorTopicYN="N">Trees</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D027843" MajorTopicYN="N">Vitis</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2008</Year><Month>7</Month><Day>12</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2009</Year><Month>1</Month><Day>10</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2008</Year><Month>7</Month><Day>12</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">18618437</ArticleId><ArticleId IdType="doi">10.1002/pca.1078</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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