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MB Sample ID: SA253929
Local Sample ID: | MSA0038 |
Subject ID: | SU002618 |
Subject Type: | Bacteria |
Subject Species: | Eggerthella lenta |
Taxonomy ID: | 84112 |
Genotype Strain: | DSM 2243 |
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Collection:
Collection ID: | CO002611 |
Collection Summary: | Time course experiments were conducted in tubes in the anaerobic chamber in a 37°C incubator. For all metabolomics experiments, three independent culture replicates were included for each condition, with an equal number of uninoculated control tubes. Starter cultures and inocula were prepared as described above for growth assays. 5mLs of defined media was added to VWR glass culture tubes (53283-800) with screw caps. The PBS-washed inoculum was added to culture tubes to obtain an approximate starting OD600 of 0.001. A preliminary growth assay was conducted to define time points spanning the exponential growth phase in the tested conditions. At each time point, OD600 measurements of all inoculated tubes were first measured using a Hach DR1900 spectrophotometer, with a paired control tube to normalize for the background. 100 μL from each tube were then transferred into a 96-well microplate, which was sealed and removed from the anaerobic chamber. Plates were centrifuged at 1,928 rcf at 4°C for 8 minutes, after which supernatants were collected into fresh polypropylene tubes or plates, sealed, and flash-frozen in liquid nitrogen. Two time course experiments were carried out with stable isotope-labeled substrates. Experimental groups included conditions in which sodium arginine in the defined media was replaced with 13C6 labeled arginine HCl, along with a matched experimental group with the same concentration of unlabeled substrate. |
Sample Type: | Bacterial culture supernatant |
Collection Frequency: | at time points specified in study design table over 56 hours (full growth phase) |
Storage Conditions: | -80℃ |