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MB Sample ID: SA330839
| Local Sample ID: | WT_C |
| Subject ID: | SU003162 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
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Sample Preparation:
| Sampleprep ID: | SP003168 |
| Sampleprep Summary: | From wild-type samples WT_A, WT_B and WT_C 5.6 x 106, 6.4 x 106 and 4.87 x 106 B lymphocytes were harvested respectively, and for knock-out samples KO_D, KO_E and KO_F 1.8 x 106, 2.1 x 106 and 1.8 x 106 B lymphocytes were harvested respectively. Cell homogenates were prepared by adding 1:1 v/v water: methanol and sonicated with a dr.hielscher UP200S (dr.hielscher, Berlin, Germany), set up to 80% intensity, 0.5 s/pulse, 16 pulses. For GC-MS, 100 µL of each homogenate was mixed with 300 µL of cold methanol containing 25 ppm D-palmitic acid (internal standard), vortex mixed for 60 min and centrifuged at 4000 g for 20 min. 250 µL of the supernatant were transferred to a vial with a glass insert and evaporated to dryness in a Gyrozen HyperVac VC2124 (Gimpo, Korea) coupled to a LVS 110Z (Gardner Denver Thomas GmbH Welch Vacuum, Fürstenfeldbruck, Germany). The samples were then submitted to methoximation (with O-methoxyamine) for 16 h and silylation for 1 h at 70ºC with N,O-Bis(trime5j,thylsilyl)trifluoroacetamide (BSTFA) and trimethylchlorosilane (TCMS), and finally resuspended in 100 µL heptane. |
| Sampleprep Protocol Filename: | Protocol_BL.pdf |
