Cells were lysed by glass beads disruption in 500 μL FA-lysis buffer (50 mmol/L HEPES-KOH at pH 7.5, 140 mmol/L NaC1, 1 mmol/L EDTA, 1% Triton X-100, 0.1% sodium deoxycholate) supplemented with 1 mmol/L PMSF and protease inhibitor cocktail (Roche).
ß-galactosidase reporter enzyme activity in the two-hybrid strains was measured using the Galacto-Star™ chemiluminescent reporter assay system according to the manufacturer's instructions (Applied Biosystems, Foster City, California), except that cells were lysed by glass bead disruption.
RNA was purified from 10 cells after glass bead disruption as described previously (Mutiu and Brandl 2005).
The cells were washed twice with sterilized DDW, and their total RNA was prepared using glass-bead disruption as described by Urushiyama et al. [39].
Total cell extracts were prepared by glass-bead disruption in lysis buffer (6 M urea, 50 mM Tris Cl pH 8.0, 0.5% SDS, 0.5% NP-40, 10 mM DDT).
Total RNA was isolated from S. cerevisiae cells by a glass-bead disruption Trizol extraction procedure and performed as described by the manufacturer (Life Technologies, Bleiswijk, The Netherlands).
P. pastoris cells containing the recombinant HBsAg protein were subjected to glass bead cell disruption.
Disruption with glass beads showed to be the most efficient method for XR release when compared to sonication process.
The hyphal mat was harvested, sequentially washed with PBS, and lysed by mechanical disruption with glass beads in a BeadBeater homogenizer (BioSpec Products, OK).
Cell extraction was performed by disruption with glass beads using a Fast Prep homogenizer.
Cell disruption using glass beads, size fractionation of mRNA-ribosome complexes on sucrose gradient and total RNA extraction were processed as described previously [ 18].
