Summary of Study ST001029

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 PR000687. The data can be accessed directly via it's Project DOI: 10.21228/M8W109 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study ID	ST001029
Study Title	An integrated, high-throughput strategy for multi-omic systems level analysis
Study Summary	This report details the automation, benchmarking, and application of a strategy for transcriptomic, proteomic, and metabolomic analyses from a common sample. The approach, Sample Preparation for multi-Omics Technologies (SPOT), provides equivalent performance to typical individual omic preparation methods, but it greatly enhances throughput and minimizes the resources required for multi-omic experiments. SPOT was applied to a multi-omics time course experiment for zinc-treated HL60 cells.
Institute	Vanderbilt University
Department	Chemistry
Last Name	Gant-Branum
First Name	Randi
Address	2201 West End Ave, Nashville, TN 37235
Email	randi.l.gant-branum@vanderbilt.edu
Phone	9312065092
Submit Date	2018-05-08
Num Groups	6
Analysis Type Detail	LC-MS
Release Date	2018-08-27
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP001075
Sampleprep Summary:	The remaining HL60 cells (approximately one million) were lysed in 100 µL of 1:1:2 MeCNCH3CN: CH3MeOH:NH4HCO3 (50 mM, pH 8.0) followed by one freeze-thaw cycle (three minutes each) and a ten-minute sonication in an ice bath. Protein concentration was obtained using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific). An aliquot of 100 or 150 µg of each sample in a total of 100 µL of lysis buffer was precipitated with 300 µL of ice cold 75:25 acetone:methanol for 2 h at -80°C. Samples were spun for 15 minutes at 6,800 x g and the supernatant was removed and utilized for metabolomics analysis. The pellets were rinsed with 300 µL of ice cold acetone and spun as above. Acetone was removed and the pellet was allowed to dry briefly. Metabolite supernatants were dried and reconstituted in 50 uL of appropriate reverse phase liquid chromatography (0.1% formic acid in 98:2 H2O: CH3CNMeCN) or hydrophilic interaction chromatography (80:20 CH3CNMeCN: H2O) compatible buffers prior to analyses.

Sampleprep ID:

SP001075

Sampleprep Summary:

The remaining HL60 cells (approximately one million) were lysed in 100 µL of 1:1:2 MeCNCH3CN: CH3MeOH:NH4HCO3 (50 mM, pH 8.0) followed by one freeze-thaw cycle (three minutes each) and a ten-minute sonication in an ice bath. Protein concentration was obtained using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific). An aliquot of 100 or 150 µg of each sample in a total of 100 µL of lysis buffer was precipitated with 300 µL of ice cold 75:25 acetone:methanol for 2 h at -80°C. Samples were spun for 15 minutes at 6,800 x g and the supernatant was removed and utilized for metabolomics analysis. The pellets were rinsed with 300 µL of ice cold acetone and spun as above. Acetone was removed and the pellet was allowed to dry briefly. Metabolite supernatants were dried and reconstituted in 50 uL of appropriate reverse phase liquid chromatography (0.1% formic acid in 98:2 H2O: CH3CNMeCN) or hydrophilic interaction chromatography (80:20 CH3CNMeCN: H2O) compatible buffers prior to analyses.