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A single colony of expression strain (GB1 or MBP) was grown overnight at 37 °C in 10 mL of filter sterilized LB media with 100% D2O substituted for water.
The swab was placed in a vial containing 1 ml of filter sterilized 1× digestion buffer [37] and stored at room temperature.
Add 10 mL of filter sterilized micronutrient solution.
TCW agar plates were prepared by mixing 25 ml of filter sterilized TCW with 25 ml of 4% agar (autoclaved) and microwaved for 30 60 seconds.
To each well, 0.5 ml of filter sterilized KM [ 37] medium containing 100 g/l mannitol, 5 μM BA, and 5 μM NAA and adjusted to pH 5.7 was added after the agarose had solidified for approximately 20 min. The cultures were maintained in the dark on an orbital shaker at 10 rpm (Belly Dancer, Stovall Life Science Inc., Greensboro, NC, USA) at a temperature of 24°C.
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Approximately 0.1 ml of filtered and non-filtered aliquots from each samples were inoculated in wells and incubated for 7 days at 40 °C under aerobic conditions.
For γ-irradiation, 140 ml of filtered HS or filtered FBS was separated into seven fractions of 20 ml each.
A solution of the LIVE/DEAD BacLight Kit L13152 (Thermo Fisher, USA) was prepared by dissolving the contents of component A and B in 5 mL of filter-sterilized ddH2O.
The filter membrane holes were cleaned with 100 mL of filtered pure water and then water samples were filtered.
To analyze the pigment ratio for each preservation period, 10 mL of floc samples were taken and poured into 100 mL of filtered sea water.
Sodium hydroxide (0.5 M) was slowly added to 10 ml of filtered wetland water while monitoring pH until a macroscopic precipitate was observed.
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