Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA202344

Local Sample ID:	M17_glu_neg_2
Subject ID:	SU002192
Subject Type:	Cultured cells
Subject Species:	Plasmodium falciparum
Taxonomy ID:	5833

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Sample Preparation:

Sampleprep ID:	SP002198
Sampleprep Summary:	Heparin synchronized Pf3D7 and PfM17-HAglmS parasites were allowed to invade RBC for 4 h and any remaining schizonts were lysed by sorbitol synchronization and parasite cultures were only treated with GlcN. Parasites were harvested at developmentally similar timepoints by centrifugation at 900 g for 5 min and then resuspended in 10 mL of chilled PBS. Parasite metabolism was quenched by cooling samples to between 3-5°C in an ethanol-dry ice bath. The rest of the preparation was performed at 4°C. Parasites were magnet purified on a VarioMACS column and 3x107 parasites were used for downstream analysis. All samples were centrifuged at 650 g for 3 min, the supernatant was removed, and the pellet washed in 500 µL of ice-cold PBS. Samples were again centrifuged at 650 g for 3 min and pellets were resuspended in 150 µL of ice-cold extraction buffer (100% methanol) and quickly resuspended. The samples were then incubated on a vortex mixer for 1 h at 4°C before being centrifuged at 17,000 g for 10 min; from this 100 µL of supernatant was collected and stored at -80°C until analysis. For each sample, another 10 µL was collected and pooled, to serve as a quality control (QC) sample.

Sampleprep ID:

SP002198

Sampleprep Summary:

Heparin synchronized Pf3D7 and PfM17-HAglmS parasites were allowed to invade RBC for 4 h and any remaining schizonts were lysed by sorbitol synchronization and parasite cultures were only treated with GlcN. Parasites were harvested at developmentally similar timepoints by centrifugation at 900 g for 5 min and then resuspended in 10 mL of chilled PBS. Parasite metabolism was quenched by cooling samples to between 3-5°C in an ethanol-dry ice bath. The rest of the preparation was performed at 4°C. Parasites were magnet purified on a VarioMACS column and 3x107 parasites were used for downstream analysis. All samples were centrifuged at 650 g for 3 min, the supernatant was removed, and the pellet washed in 500 µL of ice-cold PBS. Samples were again centrifuged at 650 g for 3 min and pellets were resuspended in 150 µL of ice-cold extraction buffer (100% methanol) and quickly resuspended. The samples were then incubated on a vortex mixer for 1 h at 4°C before being centrifuged at 17,000 g for 10 min; from this 100 µL of supernatant was collected and stored at -80°C until analysis. For each sample, another 10 µL was collected and pooled, to serve as a quality control (QC) sample.