Bio-Rad MicroRotofor™ Cell Lysis Kits User Manual
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Section 1
Introduction
MicroRotofor lysis kits provide convenient, effective methods
for the preparation of protein samples for fractionation with
the MicroRotofor cell. The MicroRotofor lysis kit (yeast) is
designed for use with yeast cultures, and employs enzymatic
digestion of the cell wall (Scott et al. 1980) followed by
solubilization into a chaotropic extraction buffer (Vuillard
et al. 1995). For added convenience, the extraction buffer
is also used as the sample buffer for isoelectric focusing
(IEF) either with the MicroRotofor cell or with IPG strips.
Section 2
Kit Specifications
Each MicroRotofor lysis kit (yeast) provides sufficient
reagent to perform at least 15 extractions (from 5 ml yeast
culture with OD
600
= 1.48, which should yield a 60 µl wet
cell pellet) and to prepare sample for 15 MicroRotofor runs.
More than 15 extractions will be possible with the kit if the
sample is applied onto IPG strips (and not prefractionated
with the MicroRotofor cell).
Each MicroRotofor run using 2.5 mg total protein yields ten
150–250 µl fractions, and the protein distribution among
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