Summary of Study ST001174

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000786. The data can be accessed directly via it's Project DOI: 10.21228/M83398 This work is supported by NIH grant, U2C- DK119886.

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Study ID	ST001174
Study Title	Role of ClpCP in respiratory and fermentative growth
Study Summary	To determine metabolite concentrations and differences at the 48 hour time point for WT, ClpC mutant, srrAB mutant, and ClpC:srrAB double mutant
Institute	Montana State University
Department	Chemistry and Biochemistry
Laboratory	Copie Lab
Last Name	Eilers
First Name	Brian
Address	103 Chemistry and Biochemistry Bldg, RM 144, Valerie Copie Lab, Bozeman, Montana, 59717, USA
Email	brian.eilers@montana.edu
Phone	4069945116
Submit Date	2019-04-24
Num Groups	4 (WT, ClpC, srrAB, and ClpC:srrAB)
Total Subjects	4
Raw Data Available	Yes
Raw Data File Type(s)	1r
Analysis Type Detail	NMR
Release Date	2019-05-15
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP001248
Sampleprep Summary:	Frozen cell pellets were resuspended in 1 mL of a 2:1 methanol/chloroform mixture and transferred to FastPrep lysis B matrix tubes (MP Biomedicals). Cells were lysed using the FastPrep-24 5G instrument and designated S. aureus settings (2 cycles at a speed of 6.0 m/s for 40 s)]. 300 μL of each layer of a 1:1 aqueous chloroform solution was added to each cell lysate. The tubes were vortexed, placed at -20 °C for 20 min, and centrifuged at 14,000 g for 10 minutes. 800 μL of the aqueous phase was transferred to microfuge tubes and placed in a Speedvac (no heat, manual run, volatile solvent) to dry overnight. Samples were resuspended in 600 μL of NMR buffer [0.25 mM 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS), 0.4 mM imidazole, 25 mM phosphate buffer, 90% H2O/10% D2O] and transferred to 5 mm Bruker NMR tubes.

Sampleprep ID:

SP001248

Sampleprep Summary:

Frozen cell pellets were resuspended in 1 mL of a 2:1 methanol/chloroform mixture and transferred to FastPrep lysis B matrix tubes (MP Biomedicals). Cells were lysed using the FastPrep-24 5G instrument and designated S. aureus settings (2 cycles at a speed of 6.0 m/s for 40 s)]. 300 μL of each layer of a 1:1 aqueous chloroform solution was added to each cell lysate. The tubes were vortexed, placed at -20 °C for 20 min, and centrifuged at 14,000 g for 10 minutes. 800 μL of the aqueous phase was transferred to microfuge tubes and placed in a Speedvac (no heat, manual run, volatile solvent) to dry overnight. Samples were resuspended in 600 μL of NMR buffer [0.25 mM 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS), 0.4 mM imidazole, 25 mM phosphate buffer, 90% H2O/10% D2O] and transferred to 5 mm Bruker NMR tubes.