Preparation of high molecular weight genomic DNA from nuclei of woody plants.
Identifieur interne : 002237 ( Ncbi/Curation ); précédent : 002236; suivant : 002238Preparation of high molecular weight genomic DNA from nuclei of woody plants.
Auteurs : F. Luro [France] ; F. LaigretSource :
- BioTechniques [ 0736-6205 ] ; 1995.
English descriptors
- KwdEn :
- MESH :
- chemical , isolation & purification : DNA, Plant.
- chemistry : Cell Nucleus.
- genetics : Plants.
- chemical , metabolism : DNA, Plant, Deoxyribonucleases, Type II Site-Specific.
- Electrophoresis, Gel, Pulsed-Field, Fruit, Molecular Weight, Nucleic Acid Hybridization.
Abstract
We have developed a rapid and reliable method for preparation of high molecular weight genomic DNA from sweet orange (Citrus sinensis) suitable for subsequent digestion by rarely cutting restriction enzymes and then separation by pulsed-field gel electrophoresis (PFGE). Methods previously described for preparation of plant DNA prior to PFGE involved protoplast isolation, a procedure that can be inefficient and time-consuming for several plant species. Nuclei isolated from plant tissues were embedded into agarose blocks and treated to release DNA, which was cleaved by restriction enzymes and then submitted to PFGE. One gram of fresh leaves gave approximately 15 micrograms of high molecular weight genomic DNA (> 2000 kbp). Within-gel hybridizations were used instead of classical Southern blotting, and the resulting signals were adequate when they were compared with those obtained with DNA prepared from crude ground leaf tissues.
PubMed: 7495551
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: Pour aller vers cette notice dans l'étape Curation :000F38
- to stream PubMed, to step Curation: Pour aller vers cette notice dans l'étape Curation :000F38
- to stream PubMed, to step Checkpoint: Pour aller vers cette notice dans l'étape Curation :000F38
- to stream Ncbi, to step Merge: Pour aller vers cette notice dans l'étape Curation :002237
Links to Exploration step
pubmed:7495551Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Preparation of high molecular weight genomic DNA from nuclei of woody plants.</title><author><name sortKey="Luro, F" sort="Luro, F" uniqKey="Luro F" first="F" last="Luro">F. Luro</name><affiliation wicri:level="1"><nlm:affiliation>Station de recherche, agronomique de Corse, INRA-CIRAD-FLHOR, San Nicolao, Villenave d'Ornon, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Station de recherche, agronomique de Corse, INRA-CIRAD-FLHOR, San Nicolao, Villenave d'Ornon</wicri:regionArea><wicri:noRegion>Villenave d'Ornon</wicri:noRegion><wicri:noRegion>Villenave d'Ornon</wicri:noRegion></affiliation></author><author><name sortKey="Laigret, F" sort="Laigret, F" uniqKey="Laigret F" first="F" last="Laigret">F. Laigret</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1995">1995</date><idno type="RBID">pubmed:7495551</idno><idno type="pmid">7495551</idno><idno type="wicri:Area/PubMed/Corpus">000F38</idno><idno type="wicri:Area/PubMed/Curation">000F38</idno><idno type="wicri:Area/PubMed/Checkpoint">000F38</idno><idno type="wicri:Area/Ncbi/Merge">002237</idno><idno type="wicri:Area/Ncbi/Curation">002237</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Preparation of high molecular weight genomic DNA from nuclei of woody plants.</title><author><name sortKey="Luro, F" sort="Luro, F" uniqKey="Luro F" first="F" last="Luro">F. Luro</name><affiliation wicri:level="1"><nlm:affiliation>Station de recherche, agronomique de Corse, INRA-CIRAD-FLHOR, San Nicolao, Villenave d'Ornon, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Station de recherche, agronomique de Corse, INRA-CIRAD-FLHOR, San Nicolao, Villenave d'Ornon</wicri:regionArea><wicri:noRegion>Villenave d'Ornon</wicri:noRegion><wicri:noRegion>Villenave d'Ornon</wicri:noRegion></affiliation></author><author><name sortKey="Laigret, F" sort="Laigret, F" uniqKey="Laigret F" first="F" last="Laigret">F. Laigret</name></author></analytic><series><title level="j">BioTechniques</title><idno type="ISSN">0736-6205</idno><imprint><date when="1995" type="published">1995</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cell Nucleus (chemistry)</term><term>DNA, Plant (isolation & purification)</term><term>DNA, Plant (metabolism)</term><term>Deoxyribonucleases, Type II Site-Specific (metabolism)</term><term>Electrophoresis, Gel, Pulsed-Field</term><term>Fruit</term><term>Molecular Weight</term><term>Nucleic Acid Hybridization</term><term>Plants (genetics)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>DNA, Plant</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Cell Nucleus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Plants</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>DNA, Plant</term><term>Deoxyribonucleases, Type II Site-Specific</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Electrophoresis, Gel, Pulsed-Field</term><term>Fruit</term><term>Molecular Weight</term><term>Nucleic Acid Hybridization</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">We have developed a rapid and reliable method for preparation of high molecular weight genomic DNA from sweet orange (Citrus sinensis) suitable for subsequent digestion by rarely cutting restriction enzymes and then separation by pulsed-field gel electrophoresis (PFGE). Methods previously described for preparation of plant DNA prior to PFGE involved protoplast isolation, a procedure that can be inefficient and time-consuming for several plant species. Nuclei isolated from plant tissues were embedded into agarose blocks and treated to release DNA, which was cleaved by restriction enzymes and then submitted to PFGE. One gram of fresh leaves gave approximately 15 micrograms of high molecular weight genomic DNA (> 2000 kbp). Within-gel hybridizations were used instead of classical Southern blotting, and the resulting signals were adequate when they were compared with those obtained with DNA prepared from crude ground leaf tissues.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Bois/explor/OrangerV1/Data/Ncbi/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002237 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Ncbi/Curation/biblio.hfd -nk 002237 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Bois
|area= OrangerV1
|flux= Ncbi
|étape= Curation
|type= RBID
|clé= pubmed:7495551
|texte= Preparation of high molecular weight genomic DNA from nuclei of woody plants.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Ncbi/Curation/RBID.i -Sk "pubmed:7495551" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Ncbi/Curation/biblio.hfd \
| NlmPubMed2Wicri -a OrangerV1
| This area was generated with Dilib version V0.6.25. |  |