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Mar 2025 17:52:24 +0100 Subject: [PATCH 4/4] fill in dead root experiment isa assay --- .../CollectionOfRootExudatesProtocol.md | 2 +- assays/DeadRootExperiment/isa.assay.xlsx | Bin 6898 -> 13635 bytes 2 files changed, 1 insertion(+), 1 deletion(-) diff --git a/assays/CollectionOfRootExudates/protocols/CollectionOfRootExudatesProtocol.md b/assays/CollectionOfRootExudates/protocols/CollectionOfRootExudatesProtocol.md index c60de24..fa8c68c 100644 --- a/assays/CollectionOfRootExudates/protocols/CollectionOfRootExudatesProtocol.md +++ b/assays/CollectionOfRootExudates/protocols/CollectionOfRootExudatesProtocol.md @@ -1,3 +1,3 @@ ## Collection of root exudates -*Arabidopsis* and *Lotus* plants were grown in a customized hydroponic system (original design by M. Peukert, University of Cologne, unpublished). This sterile growth setup consists of glass jars filled with glass beads and a stainless-steel mesh on top. Nutrient solution (modified 0.25× B&D medium, Fe-EDTA instead of Fe-citrate) was poured into the jars until the beads were covered in liquid and the liquid touched the metal mesh. We used the same medium for both plant species to allow for direct comparison of exudate composition, and to minimize differential effects on the bacterial community originating from different media types. We chose the *Lotus* B&D medium since *Arabidopsis* grew reasonably well in it. Sterilized and pregerminated seeds were placed onto the mesh, jars were put into sterile plastic boxes with filter lids (SacO2 microboxes) and plants were grown for 5 weeks. The medium containing root exudates was removed from the jars in the clean bench using a sterile metal needle and plastic syringe. After transfer to 50-ml Falcon tubes, exudates were frozen at −80 °C, freeze-dried until a volume of 2–3 ml was left, thawed and adjusted with sterile water to 5 ml. Exudates were kept at −80 °C until further usage. \ No newline at end of file +*Arabidopsis* and *Lotus* plants were grown in a customized hydroponic system (original design by M. Peukert, University of Cologne, unpublished). This sterile growth setup consists of glass jars filled with glass beads and a stainless-steel mesh on top. Nutrient solution (modified 0.25× B&D medium, Fe-EDTA instead of Fe-citrate) was poured into the jars until the beads were covered in liquid and the liquid touched the metal mesh. We used the same medium for both plant species to allow for direct comparison of exudate composition, and to minimize differential effects on the bacterial community originating from different media types. We chose the *Lotus* B&D medium since *Arabidopsis* grew reasonably well in it. Sterilized and pregerminated seeds were placed onto the mesh, jars were put into sterile plastic boxes with filter lids (SacO2 microboxes) and plants were grown for 5 weeks. The medium containing root exudates was removed from the jars in the clean bench using a sterile metal needle and plastic syringe. After transfer to 50-ml Falcon tubes, exudates were frozen at −80 °C, freeze-dried until a volume of 2–3 ml was left, thawed and adjusted with sterile water to 5 ml. 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