Rapid isolation of nuclei from carrot suspension culture cells using a BioNebulizer

The BioNebulizer is an instrument that breaks cells and large molecules using the shearing forces created by laminar flow of high-pressure gas in the microcapillary channels generated by the instrument. Within 4 min, 90% of the carrot suspension culture cells that passed through the nebulizer were b...

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Veröffentlicht in:BioTechniques. - 1988. - 16(1994), 1 vom: 02. Jan., Seite 154-9
1. Verfasser: Okpodu, C M (VerfasserIn)
Weitere Verfasser: Robertson, D, Boss, W F, Togasaki, R K, Surzycki, S J
Format: Aufsatz
Sprache:English
Veröffentlicht: 1994
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Suspensions RNA 63231-63-0
Beschreibung
Zusammenfassung:The BioNebulizer is an instrument that breaks cells and large molecules using the shearing forces created by laminar flow of high-pressure gas in the microcapillary channels generated by the instrument. Within 4 min, 90% of the carrot suspension culture cells that passed through the nebulizer were broken. Cytosol and organelles were released from the broken cells leaving cell wall ghosts. Nuclei were further purified by means of a discontinuous Percoll gradient. This method yielded an average of 2 x 10(5) nuclei from 2 g of suspension culture cells (approximately 2 x 10(6) cells). The isolated nuclei actively incorporated [8,5'-3H]-GTP into RNA
Beschreibung:Date Completed 28.04.1994
Date Revised 15.11.2006
published: Print
Citation Status MEDLINE
ISSN:0736-6205