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MB Sample ID: SA057691
Local Sample ID: | DS_33 |
Subject ID: | SU001002 |
Subject Type: | Mammal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Genotype Strain: | Fisher rat |
Age Or Age Range: | 10-week-old |
Weight Or Weight Range: | not measured |
Height Or Height Range: | not measured |
Gender: | Male |
Animal Animal Supplier: | Charles River |
Animal Housing: | standard |
Animal Light Cycle: | standard 12 h |
Animal Feed: | standard grain-based |
Animal Water: | standard unlimited |
Animal Inclusion Criteria: | random |
Species Group: | - |
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Sample Preparation:
Sampleprep ID: | SP001009 |
Sampleprep Summary: | Specimens were stored at -80ºC. Before extraction, ON and retinas were cut into ~1 mm3 pieces. 6 mL of methanol (LC-MS grade) and 3 mL of chloroform (LC-MS grade) were added to each sample. For this high throughput approach, no internal standards were added to samples. After 2 min vigorous vortexing and 2 min sonication in ultrasonic bath, the samples were incubated at 48ºC overnight (in borosilicate glass vials, PTFE-lined caps). The following day, 3 mL of water (LC-MS grade) and 1.5 mL of chloroform were added, samples vigorously vortexed for 2 min and centrifuged at 3000 RCF, 4ºC for 15 min to obtain phase separation. Lower phases were collected and dried in a centrifugal vacuum concentrator. Samples were stored at -20ºC until reconstituted in 60 µL of chloroform:methanol (1:1) prior to mass spectrometric analysis. |
Sampleprep Protocol Filename: | method.docx |
Processing Method: | lipid extraction |
Processing Storage Conditions: | Described in summary |
Extract Storage: | -20℃ |
Sample Spiking: | no internal standards added |