Summary of Study ST000321

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 PR000260. The data can be accessed directly via it's Project DOI: 10.21228/M85S3H This work is supported by NIH grant, U2C- DK119886.

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Study IDST000321
Study TitleEffects of LGG on current drinkers gut metabolism
Study SummaryThis experiment tests the effects of alcoholism by examining the primary metabolites obtained from mouse stool. Stool was collected from 4 groups of mice with varying treatments. One group was not humanized, another was humanized from a healthy human, a third was humanized from a current drinker, and the final group was humanized from a current drinker but also given a treatment of LGG. Humanizations were done to create a model of human gut activity in the mice.
Institute
University of California, Davis
DepartmentGenome and Biomedical Sciences Facility
LaboratoryWCMC Metabolomics Core
Last NameFiehn
First NameOliver
Address1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Emailofiehn@ucdavis.edu
Phone(530) 754-8258
Submit Date2014-12-22
Num Groups8
Total Subjects56
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2016-01-27
Release Version1
Oliver Fiehn Oliver Fiehn
https://dx.doi.org/10.21228/M85S3H
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP000349
Sampleprep Summary:Preparation of extraction mix and material before experiment: 1. Switch on bath to pre-cool at –20°C (2°C validity temperature range) 2. Check pH of acetonitrile and isopropanol (pH7) using wetted pH paper 3. Make the extraction solution by missing acetonitrile, isopropanol and water in proportions 3 : 3 : 2 4. Rinse the extraction solution for 5 min with nitrogen. Make sure that the nitrogen line was flushed out of air before using it for degassing the extraction solvent solution Sample Preparation 1. Weigh 50 mg tissue sample in to a 25 ml conical polypropylene centrifuge tube. 2. Add 2.5mL extraction solvent to the tissue sample and homogenize for 45 seconds ensuring that sample resembles a powder. In between samples, clean the homogenizer in solutions of methanol, acetone, water, and the extraction solvent. 3. Centrifuge the samples at 2500 rpm. for 5 minutes. Aliquot 2 X 500µl supernatant, one for analysis and one for a backup sample. Store backup aliquot in the -20°C freezer. 4. Evaporate one 500µl aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 5. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) 6. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 7. Remove supernatant to a new Eppendorff tube. 8. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 9. Submit to derivatization.
Sampleprep Protocol Filename:SOP_Extraction_of_Mammalian_Tissue_Samples.pdf
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