Sample Set Information
ID
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SE6
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Title
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Synechocystis sp. metabolite analysis
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Description
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Investigation of Synechocystis sp. PCC6803 metabolites. 3 replicates data are examined.
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Authors
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Takeshi Ara, Norimoto Shimada, Mitsuo Enomoto, Nozomu Sakurai, Hideyuki Suzuki, Daisuke Shibata, Kazusa DNA Research Institute
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Reference
|
Direct Submittion
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Comment
|
version 1
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Sample Information
ID
|
S01
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Title
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Synechocystis sp. PCC6803
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Organism - Scientific Name
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Synechocystis sp. PCC6803
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Organism - ID
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NCBI taxonomy:1148
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Compound - ID
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Compound - Source
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Preparation
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Synechocystis sp. PCC6803 are grown in the BG11 culture medium in an incubator with Light at all times condition (32 degree C, 150rpm).
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Sample Preparation Details ID
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Comment
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[KomicMarket ID] KSBA_27
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Analytical Method Details Information
ID
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MS1
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Title
|
LC-Orbitrap-MS ESI positive method 1
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Instrument
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Agilent1100 HPLC (Agilent), LTQ-Orbitrap (Thermo Fisher Scientific)
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Instrument Type
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LC-Orbitrap-MS
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Ionization
|
ESI
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Ion Mode
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Positive
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Description
|
Harvested sample is frozen by liquid N2 and resulting powder (100mg) are solved in 500uL 80% methanol solution. Cells are homogenized by TissueLyser (QUIAGEN) and centrifuged (15000rpm, 10min, 4 degree C). 20uL of the supernatant (300uL) is injected into HPLC after 0.2um membrane filter treatment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel Amide-80 (4.6 x 250 mm, 5 micrometer; TOSOH), Solvent: A; 0.1% formic acid aq. B; ACN (addition 0.1% formic acid fc.), Gradient: (B);80 to 45% (0.0 to 30.0 min), 45 to 35% (30.0 to 30.1 min), 35% (30.1 to 35.0 min), 35% to 80% (35.0 to 35.1 min), 80% (35.1 to 42.0 min), Column temp.: 40 degree C, Flow rate=0.8mL/min, PDA: 200-600 nm (2 nm step). Orbitrap-MS conditions: Filter 1: FTMS + c norm !corona !pi res=60000 o(150.0-2000.0); 2: FTMS + c norm !corona !pi res=7500 Dep MS/MS Most intense ion from (1); 3: FTMS + c norm !corona !pi res=7500 Dep MS/MS 2nd most intense ion from (1).
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Comment_of_details
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