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MB Sample ID: SA242074
Local Sample ID: | Ther-1707-02-01_35_1_1485 |
Subject ID: | SU002505 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
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Treatment:
Treatment ID: | TR002517 |
Treatment Summary: | From Normal, EAE vehicle-treated, and therapeutic SIMR1707 treated mice, the cerebellum part of the brain was cut, and immediately snap frozen using liquid nitrogen. The samples then were weighed, made in powder form and homogenized using pistol method under liquid nitrogen. The cerebellum sample was lysed using 400 uL RIBA lysis buffer and let to set for 5-10 minutes to allow lysis. 50 ul of the homogenized sample was taken for RNA extraction using norgen biotek RNA/DNA purification kit (cat#:47700) according to the manufacturer’s instructions. The remaining 350 uL was processed for metabolites and proteins extraction. First, it was centrifuged for 5 minutes at 4 C, 13000 rpm. The supernatant was collected in a new 1.5 mL eppendorf tube. The supernatant was centrifuged again for 1 minute at 4 C, at maximum speed (>13000 rpm), and the supernatant was collected again in a new 1.5 mL tube and kept on ice as it was used in the following steps, while the pellet was discarded. |