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MB Sample ID: SA201543
| Local Sample ID: | DMEM_IST_2 |
| Subject ID: | SU002189 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Genotype Strain: | OVCAR-5 and CaOV3 cell lines |
| Gender: | Not applicable |
| Cell Biosource Or Supplier: | CaOV3 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). OVCAR-5 cell line obtained from Dr. Thomas Hamilton (Fox Chase Cancer Centre, Philadelphia, PA). |
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Sample Preparation:
| Sampleprep ID: | SP002195 |
| Sampleprep Summary: | Media was aspirated and cells were washed three times with 3mL warmed PBS. Metabol-ic arrest was achieved through the addition of approximately 2 mL of liquid nitrogen di-rectly to cells ensuring that the surface of the plate was covered before plates were placed onto dry ice. Metabolite extraction was achieved through the addition of 1mL 100% ice cold methanol. Cells were lifted off of the plate using an ice cold cell scraper and transferred to a 2mL Eppendorf. An additional 1mL of 100% ice cold methanol was added be-fore snap freezing by immersion in liquid nitrogen for 3 minutes. This was followed by thawing on dry ice and vortexing to resuspend contents. Freeze/thaw process was repeat-ed 5 times to ensure full extraction of metabolites. Samples were centrifuged at 16000g at -9⁰C for 5 minutes and the supernatant was retained. The pellet was resuspended in 500uL of 100% ice cold methanol and freeze/thaw in liquid nitrogen was repeated 5 times. This sample was centrifuged at 16000g at -9⁰C for 5 minutes and the supernatant was retained and combined with previously retained supernatant. The samples were then dried in a SpeedVac Vacuum Concentrator (John Morris Scientific, RVC 2-18) at room temperature, with vacuum of 40mbar and rotor speed of 1000min-1. Before data acquisition samples were resuspended in appropriate volumes of methanol as per cell number. |
| Processing Storage Conditions: | -80℃ |
| Extraction Method: | MeOH |
| Extract Storage: | -80℃ |
| Sample Resuspension: | MeOH |
