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Culture for limonene production was carried out in 2YT medium using a shaking incubator at 20°C and 180 rpm.
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The cells were washed twice with glucose-free KRH and incubated in this medium ± 20 μM MnTMPyP for 30 min at 37 °C using a shaking plate incubator (Labnet International) set at 100 rpm.
Cells were then incubated in KRH ± 20 μM MnTMPyP (a cell-permeative superoxide dismutase mimetic [21]) for 30 min at 37 °C using a shaking plate incubator (Labnet International, Oakham, UK) set at 100 rpm.
The P. oxalicum GZ-2 strain was grown at 30°C in a 250 mL shaker flask containing 50 mL basal culture medium in a shaking incubator using 2% corncob powder as the sole carbon source.
Cultures were allowed to grow overnight at 37°C in a shaking incubator; these were used to inoculate 2 L of PASM-5052 auto-induction medium (Studier).
W2mef/AMA1-loxP/DiCre parasites were selected using blasticidin (2.5 μg ml−1, Invitrogen) and cultured in a shaking incubator to improve parasite growth post transfection (Allen and Kirk, 2010).
The flasks were incubated in a shaking incubator operated at 30 °C and 180 rpm.
The cultures were incubated in a shaking incubator maintained at 37°C and set at 220 rpm.
The culture was incubated in a shaking incubator for an hour at 30°C.
The broth cultures were incubated in a shaking incubator at 37°C for 16 - 20 hours.
This E. coli suspension was then incubated in a shaking incubator at 37 °C for 1 h.
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