MassIVE MSV000100561

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Nitrosomonas europaea Nm50 BNI degradation experiment - MBOA and D-DMPP

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Description

Nitrosomonas europaea Nm50 BNI degradation experiment - MBOA and D-DMPP
- concentrate ~ 600 ml culture
- resuspend to make mother-culture - 50 ml in 100 ml shot bottle. Feed 1 mM NH4Cl (final concentration) to mother culture bottle to keep active till 25th and setup
- Keep at 28C overnight

After two days
- Wash remaining 25 ml of mother culture (2x 12.5 in one spin column 50 ml tube: use "spent/already used 2nd tube as Tara with 12.5 ml water, during centrifugation); wash 3x with medium (check if nitrite is gone, resuspend in 170 ml medium - need 160 for setup + mother culture samples)
- let acclimate for ~20min - 40 min without substrate (lunch)
- prepare 84 ml medium with 0.5 mM ammonium for abiotic controls
- add 1 mM ammonium to mother culture, sample M0, M1 (20 min), M2 (40 min)

D-DMPP mix (790% D, 30% notz labeled) dissolved in MQ to 0.05 M (50 mM). [17.95 mg D-DMPP + 5.3 mg DMPP in 2.4 mL MQ]
Split mother culture into the following incubations, in small plastic vials I will give you:
A: 5 x 8 ml culture; mother stock 42; no DMPP (just 40 ul medium)
B: 5 X 8 ml culture mother stock 42 + 1 uM DMPP final concentration (2.1 ul emulsion)
C: 5 X 8 ml culture mother stock 42 + 2.5 uM DMPP final concentration (21 ul emulsion)
D: 5 x 8 ml culture mother stock 42 + 10 uM DMPP final concentration (210 ul emulsion)
ABIOTICS:
E: 5x 8 ml medium (with 0.5 ammonium); mother stock 42 + 1 uM DMPP final concentration (2.1 ul emulsion)
F: 5x 8 ml medium (with 0.5 ammonium) mother stock 42 + 10 uM DMPPfinal concentration (210 ul emulsion)
Take T0
Sample every 20-30-60 min; 0.5 ml for N assay and 0.5 ml for analytics (for E and F only T0, T2, T4)

Sample preparation d-DMPP samples:
1.Centrifuge samples in Eppendorf tubes at 30,000 g for 10 minutes at 4C
2.Transfer 300 uL supernatant into HPLC vials with 300/400 uL inserts

Measurements according to 10.1021/ac503290j, but with an Q Exactive HF instrument

[doi:10.25345/C51J97N43] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: biological nitrification inhibitor ; DatasetType:Metabolomics

Contact

Principal Investigators:
(in alphabetical order) 		christoph bueschl, BOKU university, Austria
Submitting User: christophb
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