Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA162666

Local Sample ID:	POL-15
Subject ID:	SU001811
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Select appropriate tab below to view additional metadata details:

Sample Preparation:

Sampleprep ID:	SP001817
Sampleprep Summary:	RNAlater solvent was removed by washing the tissue 3 times with PBS 1X. Then, the polyp was frozen in liquid nitrogen for 30 seconds. The frozen sample was put in cryoPREPTMCP02 (Covaris, MA, United States) plastic bags and submerged again for 30 seconds in liquid nitrogen. Once the plastic bag was inside the automated cryoPREPTMCP02, two consecutive impact forces of levels 2 and 4 out of 6 were applied. The resultant powder was gathered and weighted. Then, 100 µL of cold methanol:ethanol (1:1, v/v) and 0.5 µL of internal standard (LPC 18:1-d7; 0.01mM) were added per each 10 mg of tissue for metabolite extraction and protein precipitation. Samples were then vortex-mixed and homogenized using Tissue-Lyser LT homogenizer (Qiagen, Germany) for 5 min at 50 Hz, 3 times. Supernatant containing the metabolites was separated from the pellet by centrifugation (2,000 rcf for 10 min at 4 °C). Then, an aliquot of 70 µL was transferred to an LC vial and diluted with 490 µL of mobile phase (5% water: 95% acetonitrile; both with 7.5 mM ammonium acetate and 0.1% acetic acid). All samples were randomized before metabolite extraction and for the corresponding analytical run.

Sampleprep ID:

SP001817

Sampleprep Summary:

RNAlater solvent was removed by washing the tissue 3 times with PBS 1X. Then, the polyp was frozen in liquid nitrogen for 30 seconds. The frozen sample was put in cryoPREPTMCP02 (Covaris, MA, United States) plastic bags and submerged again for 30 seconds in liquid nitrogen. Once the plastic bag was inside the automated cryoPREPTMCP02, two consecutive impact forces of levels 2 and 4 out of 6 were applied. The resultant powder was gathered and weighted. Then, 100 µL of cold methanol:ethanol (1:1, v/v) and 0.5 µL of internal standard (LPC 18:1-d7; 0.01mM) were added per each 10 mg of tissue for metabolite extraction and protein precipitation. Samples were then vortex-mixed and homogenized using Tissue-Lyser LT homogenizer (Qiagen, Germany) for 5 min at 50 Hz, 3 times. Supernatant containing the metabolites was separated from the pellet by centrifugation (2,000 rcf for 10 min at 4 °C). Then, an aliquot of 70 µL was transferred to an LC vial and diluted with 490 µL of mobile phase (5% water: 95% acetonitrile; both with 7.5 mM ammonium acetate and 0.1% acetic acid). All samples were randomized before metabolite extraction and for the corresponding analytical run.