title: An improved procedure for the isolation and purification of protoplasts from carrot suspension culture.
abstract: A procedure is reported for the rapid and highly reproducible isolation of protoplasts from carrot suspension culture. The method utilizes Onozuka R 10 cellulase which has been purified by chromatography on Sephadex G75. Protoplast isolation, using this procedure, is quantitative and complete within 1 to 1.5 h. Intact protoplasts were separated from broken ones and other cellular debris by application of a polyethylene glycol/dextran two-phase system. The protoplasts isolated in this manner lack any detectable cell wall and are greater than 95% viable when assayed using fluorescein diacetate. It is concluded that such protoplasts are highly suitable for biochemical studies.
authors: Biosciences Division, Unilever Research, Colworth House, Sharnbrook, MK44 1LQ, Bedford, UK.
authors: A R
authors: AR
authors: Slabas
authors: A J
authors: AJ
authors: Powell
authors: C W
authors: CW
authors: Lloyd