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The ion association behaviour observed in our earlier studies of a polyether electrolyte system at elevated temperatures, was reminiscent of the molar conductivity behaviour typical of low dielectric constant systems.
Membrane-enriched protein fractions were prepared as previously described [18] except that cells were broken in a "One Shot" Cell Disrupter (Constant Systems LDT) at maximum pressure.
Cells were suspended in 50 mM Tris, 150 mM NaCl, 1 mM DTT (pH 7.9) and lysed using a cell disruptor (Constant Systems).
Cell disruption was performed by two passages through a cell disruptor (Constant Systems Ltd) at 39 kPsi in the presence of 1 mM MgSO4, 1 mM PMSF, 100 µg/mL DNase and 100 µg/mL RNase.
For all other preparations (including a clone expressing CifBp starting from an alternative start site, see below), cell pellets were re-suspended in 100 mM Tris, 500 mM NaCl, pH 8.2 and lysed using a Constant Systems (UK) T-series high pressure cell disruptor.
Cells were lysed using a Constant Systems cell disruptor at 20 kPSI.
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Observability analysis method named PWCS (piece-wise constant system) is applied for this new presented method.
Pellets were then resuspended in 5 ml of MeOH/CHCl3 (2:1) and mechanically disrupted twice using French Press at 28,000 psi (Constant Cell Disruption System, Constant System Ltd).
On the other hand, as cell number in the community grows with a constant system size, gene expression noise decreases.
Cells were disrupted in a One Shot Cell Disrupter (Constant System LDT, Daventry, UK) at a maximum pressure of 2.5 kbars.
Cells were harvested and lysed in lysis buffer (50 mM Tris HCl (pH 7.5), 250 mM NaCl, 1 mM PMSF, 1% Triton X-100) by means of a French Press (Constant System, Northants, UK).
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