The extracellular lipase production from E. aerogenes was carried out in 250 ml Erlenmeyer flasks each containing 50 ml of a medium composed of peptone (0.5%), yeast extract (0.3%), NaCl (0.25%), MgSO4 (0.05%) and coconut oil (3.0%) at pH 7.0.
Cells were grown in 500-mL Erlenmeyer flasks each containing 50 mL of medium and incubated at 25 °C in an orbital shaker set at 200 rpm.
Hyphae and conidia were obtained from culturing mycelia of FB, BSs, and paired BSs (Table 1) in 250 mL Erlenmeyer flasks, each of which contained a 100 mL liquid medium consisting of 3 g/L Bacto beef extract, 5 g/L Bacto peptone, and 15 g/L d-glucose, and incubated without shaking at 25 °C for 3 week.
On cooling, they were inoculated by addition of three pellets of heavily sporulating fungi from 4- to 5-day-old cultures, were picked up through cork borer (1 × 1 cm in diameter), and were added to Erlenmeyer flasks each of 250 mL capacity (10 spores mL−1, if otherwise not stated).
The media (ICCM) were harvested from the first set of flasks (directly irradiated) 1 hour postirradiation and pooled per triplicate flask at each of the time points (1, 4, 8, and 24 hours) and each of the dose points (0, 0.05, and 0.5 Gy).
Two flask of each type of cells were used every day to have precise average count of the cell.
As an additional negative control, one flask of each of the three cell lines was left untreated but monitored once a day alongside the TSV-injected flasks.
Briefly, 0.5 g samples of biochar were placed in 125-mL Erlenmeyer flasks each to which 50 mL of a 0.5 M HCl solution was added.
The scientists observed that after 500 generations, two types of E. coli were dominant in the flask, each with a distinctive set of mutations.
The three cultures were diluted to an O.D. of 0.02 in LB and grown at 37°C to an O.D. of 0.1 where they were split and 20 µg/ml of 2-AP was added to one flask of each culture.
