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MB Sample ID: SA002148
| Local Sample ID: | Heat_Treated_02 |
| Subject ID: | SU000067 |
| Subject Type: | Insect |
| Subject Species: | Centroptilum triangulifer;Atherix;Megaloptera |
| Taxonomy ID: | 2078957;124299;50553 |
| Species Group: | Insect |
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Sample Preparation:
| Sampleprep ID: | SP000063 |
| Sampleprep Summary: | Control and heat-treated mayfly samples were provided in triplicate and were processed individually. One sample was provided for AD, ControlX, MF control (from a previous study) and MG larvae. Three replicates of the larvae listed above having only a single sample were created. Aliquots of 50-300mg of insect larvae were mixed with degassed 1:1 Acetonitrile:Water solution in a 2 mL snap cap tube at a concentration of 50 mg/mL. Megalopterans were substantially larger than the other larvae and were prepared at 200 mg/mL. Samples were homogenized and then centrifuged at 4?C for 5 minutes at 14000rcf. The supernatant was removed and placed into a new tube. Samples were centrifuged again at 4?C for 5 minutes at 14000rcf and a volume of the homogenate corresponding to 40 mg insect larvae were taken. After homogenization, the three larval replicates from AD, ControlX, MG and MF control (from a previous study) were pooled and a single sample created for each larval type. There was insufficient Mayfly (heat and control) sample material to create an internal pool, so an external pool was made by combining equal concentration amounts of AD, MG and MF control (from a previous study) pool quality check (QC) samples. Samples were completely dried by vacuum centrifuge and were reconstituted by adding 630 µL of D2O (Aldrich) and 70 µL of Chenomx ISTD solution (Chenomx, Edmonton, Alberta, Canada). The samples were vortexed and centrifuged at 14000rcf for 2 minutes. 650 µL of sample was transferred into 5mm NMR tubes and analyzed by NMR. |
| Sampleprep Protocol Filename: | RTI_Metabolomic_Analysis_of_Thermally_Challenged_Larvae_Procedure.docx |
| Processing Method: | Homogenization and Solvent Removal with SpeedVac |
| Processing Storage Conditions: | 4°C |
| Extraction Method: | 50% Aqueous Acetonitrile |
| Extract Concentration Dilution: | dried on SpeedVac |
| Extract Storage: | -80C |
| Sample Resuspension: | resuspended in D2O + Chenomx ISTD solution |
