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MB Sample ID: SA120700
Local Sample ID: | WT-2 |
Subject ID: | SU001503 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Genotype Strain: | Burkitt lymphoma cell line, wild type and two mutants, 4A and VP16 |
Gender: | Not applicable |
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Sample Preparation:
Sampleprep ID: | SP001511 |
Sampleprep Summary: | The global, untargeted metabolomics study was performed on switchable MYC allele Ramos cell lines treated with 20 nM 4-OHT. Individual cell pellet samples were lysed using 200 µl ice cold lysis buffer (1:1:2, Acetonitrile : Methanol : Ammonium Bicarbonate 0.1 M, pH 8.0, LC-MS grade) and sonicated using a probe tip sonicator, 10 pulses at 30% power, cooling down on ice between samples. A BCA was used to determine the protein concentration for individual samples, and adjusted to 200 µg total protein in 200 µl of lysis buffer. Isotopically labeled standard molecules, Phenylalanine-D8 and Biotin-D2, were added to each sample to assess sample preparation. Samples were subjected to protein precipitation by addition of 800 µL of ice cold methanol (4X by volume), and incubated at -80°C overnight. Samples were centrifuged at 10,000 rpm for 10 minutes to eliminate precipitated proteins and supernatant(s) were transferred to a clean Eppendorf tube and dried down in vacuo. Samples were stored at -80°C until further LC-MS analysis. |
Sampleprep Protocol ID: | Global untargeted method_MYC_Project |
Processing Storage Conditions: | -80℃ |
Extract Storage: | -80℃ |