From 094fd9794ff62bda03e9c44e3410d6f5e0adf015 Mon Sep 17 00:00:00 2001 From: Mekhrona Mirzoeva <memir101@hhu.de> Date: Fri, 14 Mar 2025 16:06:05 +0100 Subject: [PATCH] Athaliana seedlings --- .gitattributes | 1 + assays/BacterialGrowthRates/isa.assay.xlsx | Bin 10078 -> 10079 bytes .../README.md | 0 .../dataset/.gitkeep | 0 .../isa.assay.xlsx | Bin 0 -> 14425 bytes .../protocols/.gitkeep | 0 .../R401onAthalianaSeedlingsAgarPlates.md | 3 +++ 7 files changed, 4 insertions(+) create mode 100644 assays/Mono-association experiment of R401 on Athaliana seedlings in agar plates/README.md create mode 100644 assays/Mono-association experiment of R401 on Athaliana seedlings in agar plates/dataset/.gitkeep create mode 100644 assays/Mono-association experiment of R401 on Athaliana seedlings in agar plates/isa.assay.xlsx create mode 100644 assays/Mono-association experiment of R401 on Athaliana seedlings in agar plates/protocols/.gitkeep create mode 100644 assays/Mono-association experiment of R401 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plates/protocols/R401onAthalianaSeedlingsAgarPlates.md b/assays/Mono-association experiment of R401 on Athaliana seedlings in agar plates/protocols/R401onAthalianaSeedlingsAgarPlates.md new file mode 100644 index 0000000..c7878ba --- /dev/null +++ b/assays/Mono-association experiment of R401 on Athaliana seedlings in agar plates/protocols/R401onAthalianaSeedlingsAgarPlates.md @@ -0,0 +1,3 @@ +## Mono-association experiment of R401 on A. thaliana seedlings in agar plates + +This protocol is adapted from (25). In brief, *A. thaliana* seeds were sterilized, germinated, and transferred to 1â„2 MS agar plates without sucrose, as described before. After transfer of seedlings, plants were grown for another 14 days under the same conditions. R401 wild-type and mutants were grown in 50% TSB overnight as described before. Bacterial cells were pelleted by centrifugation, washed 3x in 10 mM MgCl2 and OD~600~ was measured and adjusted to 0.0001. Agar plates were flushed with 15 ml of bacterial suspension for 5 min. The bacterial suspension were removed, and plants were carefully transferred to new 1â„2 MS agar plates. After 24 h, roots were cut using a sterile scalpel and collected in pre-weighed, sterile 2 ml tubes containing 1 steel bead (3 mm diameter). Tubes were weighed again to assess the root fresh weight. Subsequently, roots were ground in a Precellys 24 TissueLyser (Bertin Technologies) for 2 x 30 s at 6,200 rpm at 15 s intervals. Then, 250 μl of sterile 10 mM MgCl2 were added to each tube and roots were ground again under the same conditions. Each sample was subsequently 5x 1:10 diluted in sterile 10 mM MgCl2. Undiluted samples and each dilution were plated on 50% TSA square plates, dried and left to grow at 25 °C until single colonies appeared. Pictures were taken and single colonies were counted blinded. \ No newline at end of file -- GitLab