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Replicates were obtained by growing 4 independent cultures on glucose that were independently transferred to fresh media containing methanol.
The culture were resuspended into 50 mL of induction media containing methanol 0.5% (v/v) instead of glycerol to a final OD600 = 10.
For transcriptome analysis, cells were extensively pre-cultivated in batch cultures on glucose at 37°C prior to a shift to fresh media containing methanol as sole carbon source.
Methanol (0.5%) was added once a day to minimal media containing methanol to maintain the induction, for agar media this was obtained by adding 150 μL (9 cm plates) or 500 μL (14 cm plates) methanol to the lid of inverted plates.
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In a recent case that critics consider one of the most worrisome, the Methanex Corporation of Vancouver, British Columbia, is challenging California's decision to phase out the use of a gasoline additive containing methanol, which Methanex makes.
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A methanol solution (100 μL) containing methanol extracts was added to 900 μL of freshly prepared DPPH methanol solution (0.1 mM).
All strains were stored at −80 °C in either Luria Bertani (LB) broth containing 50%% glycerol (E. coli) or 50%BG-11BG-11dia containing 5%% (v/v) methanol (Synechocystis PCC6803).
Test media for feeds contained methanol extracts from feed.
Serial dilutions of the sample from the underground gas storage were prepared in liquid media containing 20 mM of methanol and 10 mM of perchlorate.
F- and C-clusters and the ULAB nerves were dissected from a cerebral ganglion preparation (for description of cerebral ganglion morphology see ref (48) and Figure 1) and separately incubated in 50 μL of extraction media containing 2 2 1 water/methanol/acetic acid overnight at 4 °C.
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