{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000968","ANALYSIS_ID":"AN001582","VERSION":"1","CREATED_ON":"May 18, 2018, 11:31 am"},

"PROJECT":{"PROJECT_TITLE":"Impact of thiamine metabolites and spent medium from Chlorella sorokiniana on metabolism in the green algae Auxenochlorella prototheciodes","PROJECT_SUMMARY":"The purpose of this study is to determine how thiamine metabolites impact central metabolism in Auxenochlorella protothecoides when grown in the presence of glucose. We hypothesize that thiamine metabolites alleviate bottlenecks in the TCA cycle and gluconeogensis, thus allowing for greater starch production when they are present. Cells were grown in bioreactors: 3 control cultures with no thiamine metabolites, 3 cultures received thiamine, 3 recieved HMP, and 3 were grown on residual medium from another algae species - Chlorella sorokiniana. We suspect that this residual medium also contains thiamine metabolites. Samples were taken daily from each of these 12 cultures over a 5 day time course so that we can observe build-up of metabolites over time.","INSTITUTE":"Auburn University","DEPARTMENT":"College of Engineering","LABORATORY":"Bioprocess Engineering","LAST_NAME":"Higgins","FIRST_NAME":"Brendan","ADDRESS":"203 Corley Building Auburn, Alabama 36849","EMAIL":"bth0023@auburn.edu","PHONE":"(334) 844-3532"},

"STUDY":{"STUDY_TITLE":"Impact of thiamine metabolites and spent medium from Chlorella sorokiniana on metabolism in the green algae Auxenochlorella prototheciodes (part I)","STUDY_SUMMARY":"The purpose of this study is to determine how thiamine metabolites impact central metabolism in Auxenochlorella protothecoides when grown in the presence of glucose. We hypothesize that thiamine metabolites alleviate bottlenecks in the TCA cycle and gluconeogensis, thus allowing for greater starch production when they are present. Cells were grown in bioreactors: 3 control cultures with no thiamine metabolites, 3 cultures received thiamine, 3 recieved HMP, and 3 were grown on residual medium from another algae species - Chlorella sorokiniana. We suspect that this residual medium also contains thiamine metabolites. Samples were taken daily from each of these 12 cultures over a 5 day time course so that we can observe build-up of metabolites over time.","INSTITUTE":"University of California, Davis","DEPARTMENT":"Genome and Biomedical Sciences Facility","LABORATORY":"WCMC Metabolomics Core","LAST_NAME":"Fiehn","FIRST_NAME":"Oliver","ADDRESS":"1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616","EMAIL":"ofiehn@ucdavis.edu","PHONE":"(530) 754-8258"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Auxenochlorella protothecoides","TAXONOMY_ID":"3075"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"D1 #1 Co Exp 2_001",
"Sample ID":"161108aOEsa16_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D1 #2 Co Exp 2_002",
"Sample ID":"161108aOEsa45_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D1 #3 Co Exp 2_003",
"Sample ID":"161108aOEsa46_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D2 #1 Co Exp 2_013",
"Sample ID":"161109aOEsa06_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D2 #2 Co Exp 2_014",
"Sample ID":"161109aOEsa01_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D2 #3 Co Exp 2_015",
"Sample ID":"161108aOEsa03_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D3 #1 Co Exp 2_025",
"Sample ID":"161108aOEsa48_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D3 #2 Co Exp 2_026",
"Sample ID":"161109aOEsa11_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D3 #3 Co Exp 2_027",
"Sample ID":"161108aOEsa31_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D4 #1 Co Exp 2_037",
"Sample ID":"161109aOEsa08_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D4 #2 Co Exp 2_038",
"Sample ID":"161108aOEsa11_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D4 #3 Co Exp 2_039",
"Sample ID":"161108aOEsa27_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D5 #1 Co Exp 2_049",
"Sample ID":"161108aOEsa21_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D5 #2 Co Exp 2_050",
"Sample ID":"161108aOEsa09_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D5 #3 Co Exp 2_051",
"Sample ID":"161109aOEsa16_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D6 #1 Co Exp 2_061",
"Sample ID":"161108aOEsa37_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D6 #2 Co Exp 2_062",
"Sample ID":"161108aOEsa40_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D6 #3 Co Exp 2_063",
"Sample ID":"161108aOEsa33_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D1 #4 Co Exp 2_004",
"Sample ID":"161109aOEsa03_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D1 #5 Co Exp 2_005",
"Sample ID":"161108aOEsa22_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D1 #6 Co Exp 2_006",
"Sample ID":"161108aOEsa14_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D2 #4 Co Exp 2_016",
"Sample ID":"161108aOEsa08_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D2 #5 Co Exp 2_017",
"Sample ID":"161108aOEsa25_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D2 #6 Co Exp 2_018",
"Sample ID":"161108aOEsa02_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D3 #4 Co Exp 2_028",
"Sample ID":"161109aOEsa21_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D3 #5 Co Exp 2_029",
"Sample ID":"161109aOEsa04_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D3 #6 Co Exp 2_030",
"Sample ID":"161108aOEsa24_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D4 #4 Co Exp 2_040",
"Sample ID":"161108aOEsa28_2",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D4 #5 Co Exp 2_041",
"Sample ID":"161108aOEsa29_2",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D4 #6 Co Exp 2_042",
"Sample ID":"161108aOEsa30_2",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D5 #4 Co Exp 2_052",
"Sample ID":"161109aOEsa05_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D5 #5 Co Exp 2_053",
"Sample ID":"161108aOEsa42_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D5 #6 Co Exp 2_054",
"Sample ID":"161108aOEsa23_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D6 #4 Co Exp 2_064",
"Sample ID":"161108aOEsa35_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D6 #5 Co Exp 2_065",
"Sample ID":"161109aOEsa14_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D6 #6 Co Exp 2_066",
"Sample ID":"161108aOEsa13_1",
"Factors":{"Treatment":"2 - Thiamine"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D1 #7 Co Exp 2_007",
"Sample ID":"161108aOEsa12_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D1 #8 Co Exp 2_008",
"Sample ID":"161109aOEsa19_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D1 #9 Co Exp 2_009",
"Sample ID":"161109aOEsa18_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D2 #7 Co Exp 2_019",
"Sample ID":"161108aOEsa47_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D2 #8 Co Exp 2_020",
"Sample ID":"161108aOEsa10_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D2 #9 Co Exp 2_021",
"Sample ID":"161108aOEsa06_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D3 #7 Co Exp 2_031",
"Sample ID":"161108aOEsa15_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D3 #8 Co Exp 2_032",
"Sample ID":"161109aOEsa13_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D3 #9 Co Exp 2_033",
"Sample ID":"161108aOEsa34_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D4 #7 Co Exp 2_043",
"Sample ID":"161108aOEsa19_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D4 #8 Co Exp 2_044",
"Sample ID":"161108aOEsa17_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D4 #9 Co Exp 2_045",
"Sample ID":"161108aOEsa18_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D5 #7 Co Exp 2_055",
"Sample ID":"161109aOEsa09_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D5 #8 Co Exp 2_056",
"Sample ID":"161109aOEsa22_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D5 #9 Co Exp 2_057",
"Sample ID":"161108aOEsa04_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D6 #7 Co Exp 2_067",
"Sample ID":"161108aOEsa44_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D6 #8 Co Exp 2_068",
"Sample ID":"161109aOEsa15_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D6 #9 Co Exp 2_069",
"Sample ID":"161109aOEsa07_1",
"Factors":{"Treatment":"3 - HMP"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D1 #10 Co Exp 2_010",
"Sample ID":"161108aOEsa43_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D1 #11 Co Exp 2_011",
"Sample ID":"161109aOEsa17_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D1 #12 Co Exp 2_012",
"Sample ID":"161108aOEsa41_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 1"}
},
{
"Subject ID":"D2 #10 Co Exp 2_022",
"Sample ID":"161108aOEsa32_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D2 #11 Co Exp 2_023",
"Sample ID":"161108aOEsa49_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D2 #12 Co Exp 2_024",
"Sample ID":"161108aOEsa39_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 2"}
},
{
"Subject ID":"D3 #10 Co Exp 2_034",
"Sample ID":"161109aOEsa10_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D3 #11 Co Exp 2_035",
"Sample ID":"161109aOEsa20_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D3 #12 Co Exp 2_036",
"Sample ID":"161108aOEsa01_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 3"}
},
{
"Subject ID":"D4 #10 Co Exp 2_046",
"Sample ID":"161108aOEsa26_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D4 #11 Co Exp 2_047",
"Sample ID":"161108aOEsa20_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D4 #12 Co Exp 2_048",
"Sample ID":"161108aOEsa38_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 4"}
},
{
"Subject ID":"D5 #10 Co Exp 2_058",
"Sample ID":"161108aOEsa05_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D5 #11 Co Exp 2_059",
"Sample ID":"161108aOEsa36_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D5 #12 Co Exp 2_060",
"Sample ID":"161109aOEsa12_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 5"}
},
{
"Subject ID":"D6 #10 Co Exp 2_070",
"Sample ID":"161109aOEsa02_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D6 #11 Co Exp 2_071",
"Sample ID":"161108aOEsa07_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
},
{
"Subject ID":"D6 #12 Co Exp 2_072",
"Sample ID":"161108aOEsa50_1",
"Factors":{"Treatment":"4 - Residual"},
"Additional sample data":{"Organ":"cell","Species":"Auxenochlorella protothecoides","Day Collected":"Day 6"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"1 ml algae cell culture was collected, 1 ml 70% MeOH (30% dH2O) at -80 C was added to the algae sample. Tubes were spun at 12,000 rcf to pellet for 2 min at 0 C. Supernatant was decanted and pellets stored at -80 before freeze drying at -45 C. Freeze dried samples were stored at -20 C until submission for extraction.","COLLECTION_PROTOCOL_FILENAME":"StudyDesignBrendanHiggins782016.pdf","SAMPLE_TYPE":"Cultured cells","VOLUMEORAMOUNT_COLLECTED":"0.15-1 mg","STORAGE_CONDITIONS":"-20℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"3 control cultures recieved no thiamine metabolites, 3 cultures received thiamine, 3 recieved HMP, and 3 were grown on residual medium from another algae species - Chlorella sorokiniana. Samples were taken daily from each of these 12 cultures over a 5 day time course so that we can observe build-up of metabolites over time.","TREATMENT_PROTOCOL_FILENAME":"StudyDesignBrendanHiggins782016.pdf"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"1. Check the pH of methanol (pH 7) 2. Make the extraction solution by mixing methanol, chloroform, and water in proportions of 10 : 3 : 1 3. Rinse the extraction solution for 5 min with nitrogen, making sure that the nitrogen line was flushed out of air before using it for degassing the extraction solvent solution. 4. Add one metal ball to each Eppendorf tube, close, and place in liquid nitrogen. 5. Take the Eppendorf tubes from the liquid nitrogen and place into the tube-holder of the grinder being careful to compensate for weight, maintaining equilibrium. 6. Shake for 30s at a frequency of 25 s-1 and check that the leaves have been ground into a fine powder. Repeat if necessary, submerging in liquid nitrogen first. 7. After grinding add 500μL of pre-chilled extraction solution to each tube one by one to prevent even partial thawing of the sample. Store all samples on ice while finishing this step. 8. Vortex the sample for 20s. 9. Centrifuge for 3min at 14,000 rcf using the centrifuge Eppendorf 5415 D. 10. Remove the whole supernatant into a clean Eppendorf tube. 11. Add 800μL of extraction solvent back into the tube with the pellet and metal ball. 12. Vortex the sample again for 20s. 13. Centrifuge for 3min at 14,000 rcf using the centrifuge Eppendorf 5415 D. 14. Remove the whole supernatant and combine with the previous supernatant. 15. Dry in the Labconco Centrivap cold trap concentrator to complete dryness and submit for derivitization.","SAMPLEPREP_PROTOCOL_FILENAME":"SOP Extraction of Plant Samples--Chlamydomonas.pdf"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Leco Pegasus III GC","COLUMN_NAME":"Restek Rtx-5Sil MS (30 x 0.25mm, 0.25um) 0.25 μm film made of 95% dimethyl/5%diphenylpolysiloxane)","FLOW_RATE":"1 ml/min","COLUMN_TEMPERATURE":"50 - 330°C","METHODS_FILENAME":"Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf","COLUMN_PRESSURE":"7.7 PSI (initial condition)","INJECTION_TEMPERATURE":"50°C ramped to 250°C by 12°C/s","SAMPLE_INJECTION":"0.5µl","OVEN_TEMPERATURE":"50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min","TRANSFERLINE_TEMPERATURE":"230°C","WASHING_BUFFER":"Ethyl Acetate","SAMPLE_LOOP_SIZE":"30 m length x 0.25 mm internal diameter","CHROMATOGRAPHY_COMMENTS":"Excel generated"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Leco Pegasus III GC TOF","INSTRUMENT_TYPE":"GC-TOF","MS_TYPE":"EI","ION_MODE":"POSITIVE","ION_SOURCE_TEMPERATURE":"250°C","IONIZATION_ENERGY":"70eV","MASS_ACCURACY":"Nominal","SOURCE_TEMPERATURE":"250°C","SCAN_RANGE_MOVERZ":"85-500","SCANNING_CYCLE":"17 Hz","SCANNING_RANGE":"80-500 Da","SKIMMER_VOLTAGE":"1850 V","MS_RESULTS_FILE":"ST000968_AN001582_Results.txt UNITS:Counts"},

"MS_METABOLITE_DATA":{
"Units":"Counts",

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}

}