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MB Sample ID: SA162630
| Local Sample ID: | control4 |
| Subject ID: | SU001808 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001808 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| control4 | SA162630 | FL019214 | Control | Factor |
Collection:
| Collection ID: | CO001801 |
| Collection Summary: | MCF7 cells were purchased from American Type Culture Collection (ATCC). All cancer cell lines were maintained in RPMI 1640 (Sigma R8758) containing glucose (2 mg/ml) and glutamine (0.6 mg/ml) but no pyruvate and was supplemented with 5% Fetal calf serum. |
| Sample Type: | Cultured cells |
Treatment:
| Treatment ID: | TR001821 |
| Treatment Summary: | MCF7 cell transfections were performed using siPORT NeoFX Transfection Agent (ThermoFisher Scientific) following the recommended protocol for an oligonucleotide siRNA based on the previously described MCJ targeting sequence. |
Sample Preparation:
| Sampleprep ID: | SP001814 |
| Sampleprep Summary: | All cancer cell lines were maintained in RPMI 1640 (Sigma R8758) containing glucose (2 mg/ml) and glutamine (0.6 mg/ml) but no pyruvate and was supplemented with 5% Fetal calf serum. Metabolic profile comparisons of cells were performed on equal cell numbers. Cells were cultured under normal conditions, detached using trypsin-EDTA (0.05 %), counted, normalized to 0.5 x 10e6 in each sample, and cell pellets were snap frozen in liquid nitrogen prior to analysis. Briefly, 2 x 10e6 cells and 20μL cell media were extracted in 1mL and 980μL cold lysis and extraction buffer (methanol : acetonitrile : water, 5:3:2), respectively, and pellets were discarded. |
Combined analysis:
| Analysis ID | AN002817 | AN002818 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Relative Abundance | Relative Abundance |
Chromatography:
| Chromatography ID: | CH002082 |
| Chromatography Summary: | Positive Mode 10 μL of water and methanol soluble fractions were run through a Kinetex C18 1.7 μm, 100 x 2.1 mm (Phenomenex) reversed phase column (Positive ion mode - phase A: water, 0.1 % formic acid; B: acetonitrile, 0.1 % formic acid) via an ultra-high performance chromatographic system (UHPLC - Vanquish, Thermo Fisher). UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in positive polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) |
| Solvent A: | 100% water; 0.1 % formic acid |
| Solvent B: | 100% acetonitrile, 0.1 % formic acid |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002083 |
| Chromatography Summary: | Negative Mode 10 μL of water and methanol soluble fractions were run through a Kinetex C18 1.7 μm, 100 x 2.1 mm (Phenomenex) reversed phase column (Negative ion mode – phase A: 1 mM NH4Ac 95:5 water : acetonitrile; phase B: 1 mM NH4Ac 95:5 acetonitrile : water) via an ultra-high performance chromatographic system (UHPLC - Vanquish, Thermo Fisher). UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in negative polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) |
| Solvent A: | 95% water/5% acetonitrile; 1 mM ammonium acetate |
| Solvent B: | 95% acetonitrile/5% water; 1 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002611 |
| Analysis ID: | AN002817 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in positive polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described. |
| Ion Mode: | POSITIVE |
| MS ID: | MS002612 |
| Analysis ID: | AN002818 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in negative polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described. |
| Ion Mode: | NEGATIVE |
