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MB Sample ID: SA094741
Local Sample ID: | CQ_E2_3 |
Subject ID: | SU001389 |
Subject Type: | Cultured cells |
Subject Species: | Plasmodium falciparum |
Taxonomy ID: | 5833 |
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Collection:
Collection ID: | CO001384 |
Collection Summary: | For metabolomics experiments, two 150 mL flasks at 6% haematocrit containing tightly synchronised ~30-34 hr trophozoites were harvested via magnet purification (Miltenyi Biotech). Infected RBC density was quantitated by flow cytometry and 2 mL of 3x 107 parasites were added to and incubated in 24 well microtiter plates for 1 hr at 37oC to stabilise the culture. Drugs (5x IC50) were added and incubated for a further 2 hrs prior to removal of the supernatant, 2x washes with 800 L ice-cold 1 x PBS with cells pelleted via centrifugation at 400 x g for 5 mins at 4oC. The cell pellets were resuspended in 150 L of ice-cold extraction buffer (MeOH) containing 1 µM internal standards; CHAPS and PIPES, and incubated on ice for 1 hr with shaking at 200 rpm. Insoluble material was pelleted with centrifugation at 14,800 x g for 10 mins at 4 oC and 120 µL of supernatant was collected and stored at -80 ºC until analysis. |
Sample Type: | Blood (whole) |