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MB Sample ID: SA272377
| Local Sample ID: | Sample_19 |
| Subject ID: | SU002814 |
| Subject Type: | Cultured cells |
| Subject Species: | NCCFH1, UOK262, UTFHC1 |
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Collection:
| Collection ID: | CO002807 |
| Collection Summary: | Place the 6 well plates to be extracted on ice. Completely aspirate off the media from each well. Gently add 2mL of ice cold ammonium acetate to each and every well without disturbing the cells. Aspirate off the ammonium acetate. Repeat this wash step once more. Aspirate off as much ammonium acetate as possible. Add 500uL of the 80% methanol solution to each and every well. Place all the 6 well plates in a -80°C freezer for 15 min. Remove the plates from the freezer, place them back on ice, and use cell scrapers to scrape off the adherent cells into solution. Transfer the cell solution from each well into a new eppitube. Vortex all the eppitubes vigorously. Centrifuge the eppitubes at 17,000g for 10 min at 4°C. Transfer the top 250 uL of the supernatant into a new 2mL tube for evaporation. Move all the tubes to the N2 evaporator and open them all. Then adjust all the needles to point into the center of the 2mL tubes and lower them appropriately (slightly above the rim of the tube) to ensure efficient evaporation. Make sure all needles are open by turning the control valve above them to the on position with the red arrow pointing down. Very slowly and carefully open the nitrogen tank to allow gas to flow into the needles. Use the pressure gauge adjacent to the evaporator to make sure gas flow is in the appropriate range. Once the drying begins it usually takes about an hour to finish. Remove the tubes from the N2 evaporator post drying and individually inspect them to ensure that all the solvent has evaporated. Places the dried samples in the -80°C freezer for storage until they can be run. |
| Sample Type: | Cultured cells |
| Storage Conditions: | -80℃ |
