{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000969","ANALYSIS_ID":"AN001583","VERSION":"1","CREATED_ON":"May 18, 2018, 11:41 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 II)","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 3_001",
"Sample ID":"161115aZKsa04_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D1 #2 Co Exp 3_002",
"Sample ID":"161116aZKsa17_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D1 #3 Co Exp 3_003",
"Sample ID":"161115aZKsa08_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D2 #1 Co Exp 3_013",
"Sample ID":"161115aZKsa39_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D2 #2 Co Exp 3_014",
"Sample ID":"161115aZKsa49_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D2 #3 Co Exp 3_015",
"Sample ID":"161115aZKsa28_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D3 #1 Co Exp 3_025",
"Sample ID":"161116aZKsa03_2",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D3 #2 Co Exp 3_026",
"Sample ID":"161115aZKsa13_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D3 #3 Co Exp 3_027",
"Sample ID":"161115aZKsa31_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D4 #1 Co Exp 3_037",
"Sample ID":"161115aZKsa17_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D4 #2 Co Exp 3_038",
"Sample ID":"161116aZKsa14_2",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D4 #3 Co Exp 3_039",
"Sample ID":"161116aZKsa22_2",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D5 #1 Co Exp 3_049",
"Sample ID":"161115aZKsa43_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D5 #2 Co Exp 3_050",
"Sample ID":"161115aZKsa07_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D5 #3 Co Exp 3_051",
"Sample ID":"161115aZKsa47_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D6 #1 Co Exp 3_061",
"Sample ID":"161116aZKsa08_2",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D6 #2 Co Exp 3_062",
"Sample ID":"161115aZKsa10_1",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D6 #3 Co Exp 3_063",
"Sample ID":"161116aZKsa06_2",
"Factors":{"Treatment":"1 - Control"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D1 #7 Co Exp 3_007",
"Sample ID":"161116aZKsa12_3",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D1 #8 Co Exp 3_008",
"Sample ID":"161115aZKsa15_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D1 #9 Co Exp 3_009",
"Sample ID":"161115aZKsa03_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D2 #7 Co Exp 3_019",
"Sample ID":"161115aZKsa05_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D2 #8 Co Exp 3_020",
"Sample ID":"161115aZKsa12_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D2 #9 Co Exp 3_021",
"Sample ID":"161115aZKsa01_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D3 #7 Co Exp 3_031",
"Sample ID":"161115aZKsa50_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D3 #8 Co Exp 3_032",
"Sample ID":"161115aZKsa27_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D3 #9 Co Exp 3_033",
"Sample ID":"161116aZKsa10_2",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D4 #7 Co Exp 3_043",
"Sample ID":"161116aZKsa02_2",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D4 #8 Co Exp 3_044",
"Sample ID":"161115aZKsa40_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D4 #9 Co Exp 3_045",
"Sample ID":"161116aZKsa05_2",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D5 #7 Co Exp 3_055",
"Sample ID":"161115aZKsa22_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D5 #8 Co Exp 3_056",
"Sample ID":"161115aZKsa25_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D5 #9 Co Exp 3_057",
"Sample ID":"161115aZKsa24_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D6 #7 Co Exp 3_067",
"Sample ID":"161115aZKsa29_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D6 #8 Co Exp 3_068",
"Sample ID":"161116aZKsa16_1",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D6 #9 Co Exp 3_069",
"Sample ID":"161116aZKsa09_2",
"Factors":{"Treatment":"2 - HMP"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D1 #10 Co Exp 3_010",
"Sample ID":"161116aZKsa01_2",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D1 #11 Co Exp 3_011",
"Sample ID":"161115aZKsa36_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D1 #12 Co Exp 3_012",
"Sample ID":"161116aZKsa07_2",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D2 #10 Co Exp 3_022",
"Sample ID":"161115aZKsa30_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D2 #11 Co Exp 3_023",
"Sample ID":"161116aZKsa20_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D2 #12 Co Exp 3_024",
"Sample ID":"161115aZKsa26_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D3 #10 Co Exp 3_034",
"Sample ID":"161115aZKsa46_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D3 #11 Co Exp 3_035",
"Sample ID":"161115aZKsa11_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D3 #12 Co Exp 3_036",
"Sample ID":"161115aZKsa37_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D4 #10 Co Exp 3_046",
"Sample ID":"161116aZKsa11_3",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D4 #11 Co Exp 3_047",
"Sample ID":"161115aZKsa02_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D4 #12 Co Exp 3_048",
"Sample ID":"161115aZKsa45_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D5 #10 Co Exp 3_058",
"Sample ID":"161116aZKsa04_2",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D5 #11 Co Exp 3_059",
"Sample ID":"161116aZKsa21_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D5 #12 Co Exp 3_060",
"Sample ID":"161115aZKsa19_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D6 #10 Co Exp 3_070",
"Sample ID":"161115aZKsa41_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D6 #11 Co Exp 3_071",
"Sample ID":"161115aZKsa23_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D6 #12 Co Exp 3_072",
"Sample ID":"161115aZKsa32_1",
"Factors":{"Treatment":"3 - Residual"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D1 #4 Co Exp 3_004",
"Sample ID":"161115aZKsa44_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D1 #5 Co Exp 3_005",
"Sample ID":"161115aZKsa18_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D1 #6 Co Exp 3_006",
"Sample ID":"161115aZKsa42_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 1"}
},
{
"Subject ID":"D2 #4 Co Exp 3_016",
"Sample ID":"161115aZKsa14_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D2 #5 Co Exp 3_017",
"Sample ID":"161115aZKsa16_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D2 #6 Co Exp 3_018",
"Sample ID":"161115aZKsa33_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 2"}
},
{
"Subject ID":"D3 #4 Co Exp 3_028",
"Sample ID":"161116aZKsa15_2",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D3 #5 Co Exp 3_029",
"Sample ID":"161115aZKsa06_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D3 #6 Co Exp 3_030",
"Sample ID":"161115aZKsa21_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 3"}
},
{
"Subject ID":"D4 #4 Co Exp 3_040",
"Sample ID":"161116aZKsa19_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D4 #5 Co Exp 3_041",
"Sample ID":"161115aZKsa20_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D4 #6 Co Exp 3_042",
"Sample ID":"161115aZKsa38_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 4"}
},
{
"Subject ID":"D5 #4 Co Exp 3_052",
"Sample ID":"161115aZKsa09_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D5 #5 Co Exp 3_053",
"Sample ID":"161116aZKsa18_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D5 #6 Co Exp 3_054",
"Sample ID":"161116aZKsa13_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 5"}
},
{
"Subject ID":"D6 #4 Co Exp 3_064",
"Sample ID":"161115aZKsa35_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D6 #5 Co Exp 3_065",
"Sample ID":"161115aZKsa48_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"Day 6"}
},
{
"Subject ID":"D6 #6 Co Exp 3_066",
"Sample ID":"161115aZKsa34_1",
"Factors":{"Treatment":"4 - Thiamine"},
"Additional sample data":{"Species":"Auxenochlorella protothecoides","Organ":"cell","Collection Day":"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":"StudyDesignBrendanHiggins7252016.pdf","SAMPLE_TYPE":"Cultured cells","VOLUMEORAMOUNT_COLLECTED":"0.11-3 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":"StudyDesignBrendanHiggins7252016.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":"ST000969_AN001583_Results.txt UNITS:Counts"},

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

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