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MB Sample ID: SA235670
Local Sample ID: | JM SQ 2 |
Subject ID: | SU002437 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
Genotype Strain: | BW25113 |
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Sample Preparation:
Sampleprep ID: | SP002443 |
Sampleprep Summary: | Cell pellets were resuspended in chilled extraction solution (500 µL) comprised of 3:1 MeOH: H2O and (13C5, 15N)valine (1 mM, 0.5 µL) and (13C6)sorbitol (1 mM, 0.5 µL) as internal standards. The suspensions were subjected to freeze-thaw cycles to facilitate lysis of the cells (30 s in liquid N2, 30 s in a dry ice/EtOH bath for 10 cycles), and shaken (9000 rpm, 10 min, 2°C). The samples were then centrifuged (12700 rpm, 5 min, 1°C). The cell lysate supernatant was transferred into glass inserts and dried. All samples were washed with MeOH (50 µL). All samples were methoximated with methoxylamine hydrochloride solution (30 mg/mL in pyridine, 20 µL) for 2 h at 37°C, followed by trimethylsilyation in BSTFA + 1% TMCS (20 µL) for 1 h at 37°C with continuous mixing. Samples were incubated at rt for 1 h prior to GC-MS analysis. |