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Additionally, the original 70 mM buffer system was replaced by a 10 mM buffer system when B. subtilis was cultivated in a benchtop bioreactor allowing pH control.
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The samples were incubated at 70°C for 15 minutes, and then chilled to room temperature, neutralized with 4.5 μl 1 M HCl and purified using the MinElute Reaction Cleanup system (Qiagen), following the manufacturer's recommendations, except that the wash and elution buffers provided with the system were replaced by 80% ethanol and 100 mM NaHCO3 (pH 9.0), respectively.
Cisco Systems is replacing GM, and Travelers Cos. replaces Citi.
For buffer desalting, the ribosome dilution buffer was replaced with PBS through ultrafiltration (Amicon Ultra 3000 Da MW; Merck, Darmstadt, Germany).
The uptake was also measured in sodium-free incubation buffer (Na+ was replaced with N-methylglucamine).
In Na+ free uptake buffer, NaCl was replaced with 96 mM choline chloride.
On each occasion the collected buffer was replaced with fresh buffer to maintain constant volume.
After that, the buffer was replaced to TE buffer with no supplement and stored at 4 °C.
The buffer was replaced with a fresh buffer containing 3.7 mM glucose for one hour before the buffer were collected (basal) and replaced with a buffer containing 16.7 mM glucose for an additional one hour (glucose stimulated).
After the formation of GUVs, the outer buffer was replaced by a fresh buffer-1 containing 200 mM glucose.
Alternatively, rehydration buffer was replaced by 0.5% IPG buffer in distilled water for native IEF.
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