OrangerV1, France, Analysis, bibRecord, 000244

Preparation of high molecular weight genomic DNA from nuclei of woody plants

Identifieur interne : 000244 ( France/Analysis ); précédent : 000243; suivant : 000245

Preparation of high molecular weight genomic DNA from nuclei of woody plants

Auteurs : F. Luro [France] ; F. Laigret [France]

Source :

BioTechniques [ 0736-6205 ] ; 1995.

RBID : Pascal:95-0518614

Descripteurs français

Pascal (Inist)
- Electrophorèse champ pulsé, Hydrolyse, Purification, DNA, Génome, Citrus sinensis, Méthode Southern, Restriction endonuclease, Feuille végétal, Agrume.
Wicri :
- topic : Agrume.

English descriptors

KwdEn :
- Cell Nucleus (chemistry), Citrus fruit, Citrus sinensis, DNA, DNA, Plant (isolation & purification), DNA, Plant (metabolism), Deoxyribonucleases, Type II Site-Specific (metabolism), Electrophoresis, Gel, Pulsed-Field, Fruit, Genome, Hydrolysis, Molecular Weight, Nucleic Acid Hybridization, Plant leaf, Plants (genetics), Pulsed field electrophoresis, Purification, Restriction endonuclease, Southern blotting.
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 μg 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.

Affiliations:

Links to Exploration step

Pascal:95-0518614

Le document en format XML

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<term>DNA, Plant (metabolism)</term>
<term>Deoxyribonucleases, Type II Site-Specific (metabolism)</term>
<term>Electrophoresis, Gel, Pulsed-Field</term>
<term>Fruit</term>
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<term>Hydrolysis</term>
<term>Molecular Weight</term>
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<term>Molecular Weight</term>
<term>Nucleic Acid Hybridization</term>
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<term>Méthode Southern</term>
<term>Restriction endonuclease</term>
<term>Feuille végétal</term>
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<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 μg 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>
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Preparation of high molecular weight genomic DNA from nuclei of woody plants

Preparation of high molecular weight genomic DNA from nuclei of woody plants

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Links to Exploration step

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

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