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MB Sample ID: SA162728
Local Sample ID: | CUcovID_0092 |
Subject ID: | SU001812 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
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Sample Preparation:
Sampleprep ID: | SP001818 |
Sampleprep Summary: | Samples were thawed on ice and extracted via a modified Folch method (chloroform/methanol/water 8:4:3), which completely inactivates other coronaviruses, such as MERS-CoV. Briefly, 20 μL of sample was diluted in 130 μL of LC-MS grade water, 600 μL of ice-cold chloroform/methanol (2:1) was added, and the samples were vortexed for 10 seconds. Samples were then incubated at 4°C for 5 minutes, quickly vortexed (5 seconds), and centrifuged at 14,000 g for 10 minutes at 4°C. The top (i.e., aqueous) phase was transferred to a new tube for metabolomics analysis and flash frozen. The bottom (i.e., organic) phase was transferred to a new tube for lipidomics analysis, then dried under N2 flow. UHPLC-MS metabolomics. Analyses were performed using a Vanquish UHPLC coupled online to a Q Exactive high resolution mass spectrometer (Thermo Fisher Scientific, Bremen, Germany). Samples (10 uL per injection) were randomized and analyzed in positive and negative electrospray ionization modes (separate runs) using a 5-minute C18 gradient on a Kinetex C18 column (Phenomenex) as described (Nemkov et al., 2019). Data were analyzed using Maven (Princeton University, Princeton, NJ, USA) in conjunction with the KEGG database and an in-house standard library. |
Processing Storage Conditions: | 4℃ |
Extract Storage: | -80℃ |