Manual

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 (bacteria) is
designed for use with both gram negative and gram positive
bacterial cultures, and employs enzymatic digestion of the
cell wall (Repaske et al. 1956) 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 kit provides sufficient reagent to prepare 15 extractions
for loading onto the MicroRotofor. This is based on each
lysis being performed on 4.5 ml fresh E. coli culture (OD
600
=
1.455), which should yield a wet cell pellet of roughly 50 µl.
More than 15 extractions will be possible if the sample is
applied onto IPG strips (and not prefractionated with the
MicroRotor cell).
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