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Okpodu
CM, Robertson D, Boss WF, Togasaki RK, Surzycki SJ (1994) Biotechniques
16: 154-159
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 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.
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