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Artificial seawater medium modified to contain 5 mM calcium was used for diatoms (ASP2,[25]) and marine broth, 2216 [Difco] for bacteria.
In the present study, we exposed fibroblast-seeded three-dimensional collagen chitosan hydrogels to a defined culture medium modified to have specific concentrations of ions involved in biomineralization.
To monitor growth in Middlebrook 7H9 and Sauton's medium (with 0.2% glycerol) or Sauton's medium modified to contain 0.1% butyrate as carbon source, 30 ml of the different media were inoculated with aliquots of the different input mixes at an OD580 of 0.01.
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Based on this finding, the culture medium was modified to increase intracellular ATP concentration by cell growth inhibition.
After reaching confluence, the medium was modified to include ascorbic acid at 50-μg/ml [ 51].
Cells were cultured in 1 L flasks filled with 750 ml of Erdschreiber's medium (UTEX), modified to have different concentrations of nitrogen and salinity.
When the M medium is modified to contain 7.5 mM ammonium chloride or 7.5 mM potassium nitrate, the amount of THB1 within C. reinhardtii strain CC-1690 varies.
Following routine sub-passaging, 3.5×10 mouse ESCs were transferred to 30 mL differentiation medium (ESM modified to contain 10% ESC-qualified fetal calf serum and without LIF) in a 10 cm ultra-low attachment suspension culture dish (Corning, Lowell, MA).
Afterwards, each type of bacterial lawn was prepared by spreading 25 μL of the bacterial suspension in M9 buffer, corresponding to 10 mg of bacterial cells, on nematode growth medium (NGM) modified to be peptone-free (mNGM) in 3.5 cm diameter plates, according to Ikeda et al. (2007) [ 19].
Strains of the in-house strain collection, exhibiting cyclic amidase activity, were cultivated in LB medium modified according to Bertani (1951) (10.0 g/L tryptone, 5.0 g/L yeast extract, 10.0 g/L NaCl, adjusted to pH 7.0).
Phlebotomine sand flies were mounted permanently on slides using Euparal or Canada balsam, after clearing in 20% potassium for two hours, washing in distilled water twice for 30 minutes, clearing in Hoyer medium modified according to Marc-André for 1 hour, dehydratation in 70% (30 minutes) and then 90% alcohol (30 minutes).
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