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In Figure 8, the FR LED source was filtered with two layers of Italian Blue gel (Roscolux; Roscolab Ltd, Sydenham UK), resulting in a λmax of 760nm.\n\nHalf the samples were entrained to inverted LD cycles, so that harvesting of day and night samples fell within a 12h interval on each of the six days of sampling.", "temperature_conditions": { "label": "const. 22.0C", "env_conditions": { "type": "constant_temp", "label": "const. 22.0C", "duration": "4", "env_day": { "type": "constant_temp", "label": "constant temperature", "duration": "4", "cycle_length": "24", "base_value": "22.0", "second_value": "22.0" } } }, "light_conditions": { "label": "F: LD 12:00:12:00", "light_channel": { "spectrum": "far red", "intensity": "2", "source": "LED", "env_conditions": { "type": "diurnal_light", "label": "LD 12:00:12:00", "duration": "4", "env_day": { "type": "diurnal_light", "label": "diurnal light", "duration": "4", "cycle_length": "24", "base_value": "0.0", "second_value": "100.0", "env_period": { "start_time": "0:00", "duration": "12:00" } } } } } }, "experimental_condition": { "name": "1 x FR-D - 3 x cFR - 2 x DD", "medium": "Agar 2% MS 1 3% Sucrose", "growth_space": "Binder growth cabinet", "duration": "6.0", "comment": "Half the samples were entrained to inverted LD cycles, so that harvesting of day and night samples fell within a 12h interval on each of the six days of sampling.", "temperature_conditions": { "label": "const. 22.0C", "env_conditions": { "type": "constant_temp", "label": "const. 22.0C", "duration": "6", "env_day": { "type": "constant_temp", "label": "constant temperature", "duration": "6", "cycle_length": "24", "base_value": "22.0", "second_value": "22.0" } } }, "light_conditions": { "label": "F: 1*LD 12:00:12:00; 3*LL; 2*DD", "light_channel": { "spectrum": "far red", "intensity": "2", "source": "LED", "env_conditions": { "type": "light_complex", "label": "1*LD 12:00:12:00; 3*LL; 2*DD", "duration": "6", "env_day": [ { "type": "diurnal_light", "label": "diurnal light", "duration": "1", "cycle_length": "24", "base_value": "0.0", "second_value": "100.0", "env_period": { "start_time": "0:00", "duration": "12:00" } }, { "type": "constant_light", "label": "constant light", "duration": "3", "cycle_length": "24", "base_value": "0.0", "second_value": "100.0", "env_period": { "start_time": "0:00", "duration": "24:00" } }, { "type": "constant_dark", "label": "constant dark", "duration": "2", "cycle_length": "24", "base_value": "0.0", "second_value": "0.0" } ] } } } }, "measurement": { "technique": "RT-PCR", "equipment": "Roche LightCycler 480", "parameters": "The qPCR was set up with a liquid handling robot\n(TECAN freedom Evo, http://www.tecan.com/) and cDNAs for each\nsample were quantified in triplicate using the LightCycler 480 Real-\nTime PCR System (Roche, http//http://www.roche.com/). 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The two comparisons are uploaded as separate experiments.", "pubmed_id": "21255161", "provenance": { "pub_title": "Light inputs shape the Arabidopsis circadian system", "first_author": "Benedicte Wenden", "group": "Millar", "pub_date": "2011-03-04" }, "growth_condition": { "name": "FR-D cycles", "medium": "Agar 2% MS 1 3% Sucrose", "stratification": "4 days at 4C; 3h of 80 μmol m-2 s-1 of fluorescent white light at 22°C were applied to allow germination., then 21h of darkness at 22°C.", "growth_space": "Binder growth cabinet", "duration": "4", "comment": "The seeds were stratified at 4°C for 96h in darkness. 3h of 80 μmol m-2 s-1 of fluorescent white light at 22°C were applied to allow germination. After 21h of darkness at 22°C, seeds were entrained under (12 hr/12 hr) light/dark cycles of 2 μmol m−2 sec−1 FR (Figure 8 of the TPJ paper). In Figure 8, the FR LED source was filtered with two layers of Italian Blue gel (Roscolux; Roscolab Ltd, Sydenham UK), resulting in a λmax of 760nm.\n\nHalf the samples were entrained to inverted LD cycles, so that harvesting of day and night samples fell within a 12h interval on each of the six days of sampling.", "temperature_conditions": { "constant_temp": { "cycle_length": "24", "base_temp": "22" } }, "light_conditions": { "light_channel": { "spectrum": "far red", "intensity": "2", "source": "LED", "diurnal_light": { "cycle_length": "24", "start_time": "0", "duration": "12" } } } }, "experimental_condition": { "name": "1 x FR-D - 3 x cFR - 2 x DD", "medium": "Agar 2% MS 1 3% Sucrose", "growth_space": "Binder growth cabinet", "duration": "6", "comment": "Half the samples were entrained to inverted LD cycles, so that harvesting of day and night samples fell within a 12h interval on each of the six days of sampling.", "temperature_conditions": { "constant_temp": { "cycle_length": "24", "base_temp": "22" } }, "light_conditions": { "light_channel": { "spectrum": "far red", "intensity": "2", "source": "LED", "light_complex_settings": { "light_days": [ { "start_day": "1", "duration": "1", "diurnal_light": { "cycle_length": "24", "start_time": "0", "duration": "12" } }, { "start_day": "2", "duration": "3", "constant_light": { "cycle_length": "24" } }, { "start_day": "5", "duration": "2", "constant_dark": { "cycle_length": "24" } } ] } } } }, "measurement": { "technique": "RT-PCR", "equipment": "Roche LightCycler 480", "parameters": "The qPCR was set up with a liquid handling robot\n(TECAN freedom Evo, http://www.tecan.com/) and cDNAs for each\nsample were quantified in triplicate using the LightCycler 480 Real-\nTime PCR System (Roche, http//http://www.roche.com/). Primers\nused are described in the Supporting Information." }, "sample_preparation": { "method": "Approximately 75–100 seedlings were harvested in RNAlater\n(Sigma, http://www.sigmaaldrich.com/) and total RNA was\nextracted using a Plant RNeasy kit (Qiagen, http://www.qiagen.com/)\nwith on-column DNase digestion. The cDNA samples for real-time\nPCR applications were reverse transcribed from 1 lg of RNA using\nthe SuperScript VILO cDNA Synthesis kit (Invitrogen, http://\nwww.invitrogen.com/), and the cDNA product was diluted 1:10 in\nRNase-free water." }, "samples": { "sample": [ { "sample_id": "1", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "Wild Type" }, "marker": "CCA1", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" }, { "sample_id": "2", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "elf4-101" }, "marker": "CCA1", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" }, { "sample_id": "3", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "Wild Type" }, "marker": "LHY", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" }, { "sample_id": "4", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "elf4-101" }, "marker": "LHY", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" }, { "sample_id": "5", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "Wild Type" }, "marker": "PRR9", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" }, { "sample_id": "6", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "elf4-101" }, "marker": "PRR9", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" }, { "sample_id": "7", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "Wild Type" }, "marker": "GI", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" }, { "sample_id": "8", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "elf4-101" }, "marker": "GI", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" }, { "sample_id": "9", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "Wild Type" }, "marker": "TOC1", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" }, { "sample_id": "10", "sample_type": "seedling", "origin": "seedling", "growth_stage": "seedling", "genetic_info": { "species": "Arabidopsis thaliana", "background": "Columbia (Col)", "genotype": "elf4-101" }, "marker": "TOC1", "growth_cond_name": "FR-D cycles", "experiment_cond_name": "1 x FR-D - 3 x cFR - 2 x DD" } ] } }, "biblio": { "principal": "Andrew Millar", "institution": "University of Edinburgh", "author": "Benedicte Wenden" } }