From f30198a359c039bd032f07f14b9df2b066519b34 Mon Sep 17 00:00:00 2001 From: Mekhrona Mirzoeva <memir101@hhu.de> Date: Thu, 6 Feb 2025 16:01:02 +0100 Subject: [PATCH] metabolomics update and DAPGdetection assay --- .../README.md | 0 .../dataset/.gitkeep | 0 .../isa.assay.xlsx | Bin 0 -> 11885 bytes .../protocols/.gitkeep | 0 .../protocols/DAPGandPyoverdineDetection.md | 3 +++ assays/MetabolomicAnalysis/isa.assay.xlsx | Bin 10742 -> 10742 bytes .../isa.assay.xlsx | Bin 11666 -> 11666 bytes isa.investigation.xlsx | Bin 7876 -> 7876 bytes .../isa.study.xlsx | Bin 12308 -> 12308 bytes .../Plant Growth Conditions/isa.study.xlsx | Bin 15939 -> 15939 bytes 10 files changed, 3 insertions(+) create mode 100644 assays/DetectionOfR401DAPGandPyoverdine/README.md create mode 100644 assays/DetectionOfR401DAPGandPyoverdine/dataset/.gitkeep create mode 100644 assays/DetectionOfR401DAPGandPyoverdine/isa.assay.xlsx create mode 100644 assays/DetectionOfR401DAPGandPyoverdine/protocols/.gitkeep create mode 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a/assays/DetectionOfR401DAPGandPyoverdine/protocols/DAPGandPyoverdineDetection.md b/assays/DetectionOfR401DAPGandPyoverdine/protocols/DAPGandPyoverdineDetection.md new file mode 100644 index 0000000..b81ba3a --- /dev/null +++ b/assays/DetectionOfR401DAPGandPyoverdine/protocols/DAPGandPyoverdineDetection.md @@ -0,0 +1,3 @@ +## Detection of R401 DAPG and pyoverdine + +For the detection of iron-chelating compounds, a R401 preculture was grown over night in 5 ml LB medium at 30 °C and 200 rpm. Before inoculation of the main culture, cells were washed twice with the main culture medium to remove potential traces of iron from the medium. Erlenmeyer flasks containing 20 ml of modified MM63 (KH2PO4 13.61 g/L, KOH 4.21 g/L, (NH4)SO4 1.98 g/L, MgSO4*7 H2O 0.25 g/L, NaCl 0.5g/L, Glucose * H2O 5,00 g/L, pH 7.1 KOH/HCl) with or without addition of 0.0011 g/L FeSO4 * 7 H2O were inoculated with 100 μl of preculture and cultivated for 6 days at 30 °C and 200 rpm. Every second day, 0.5 ml sample were taken, cells were removed by centrifugation and 5 μl of supernatant were analysed on a Bruker microTOFq-II high-resolution mass spectrometer coupled to an Agilent 1290 UPLC system with an Acquity UPLC BEH C-18 reverse phase column, run in a gradient of MeCN/H2O + 0.1% formic acid. 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