From 0814146b9b2df3548bfb13ac94c7ea58848109ed Mon Sep 17 00:00:00 2001
From: Dominik Brilhaus <brilhaus@nfdi4plants.org>
Date: Mon, 8 Jan 2024 14:40:50 +0100
Subject: [PATCH] restructure

---
 .../isa.assay.xlsx                            | Bin 18247 -> 12013 bytes
 .../isa.assay.xlsx                            | Bin 2684 -> 6938 bytes
 assays/2_2-FAME-analysis/README.md            |   0
 assays/2_2-FAME-analysis/dataset/.gitkeep     |   0
 assays/2_2-FAME-analysis/isa.assay.xlsx       | Bin 0 -> 6901 bytes
 assays/2_2-FAME-analysis/protocols/.gitkeep   |   0
 .../protocols/2_2-FAME-analysis.md            |   8 ++++++++
 isa.investigation.xlsx                        | Bin 8306 -> 8306 bytes
 studies/Ricinus-communis-seeds/README.md      |   0
 studies/Ricinus-communis-seeds/isa.study.xlsx | Bin 0 -> 7537 bytes
 .../Ricinus-communis-seeds/protocols/.gitkeep |   0
 .../protocols/Plant-growth-conditions.md      |   6 ++++++
 .../Ricinus-communis-seeds/resources/.gitkeep |   0
 workflows/.gitkeep                            |   0
 14 files changed, 14 insertions(+)
 create mode 100644 assays/2_2-FAME-analysis/README.md
 create mode 100644 assays/2_2-FAME-analysis/dataset/.gitkeep
 create mode 100644 assays/2_2-FAME-analysis/isa.assay.xlsx
 create mode 100644 assays/2_2-FAME-analysis/protocols/.gitkeep
 create mode 100644 assays/2_2-FAME-analysis/protocols/2_2-FAME-analysis.md
 create mode 100644 studies/Ricinus-communis-seeds/README.md
 create mode 100644 studies/Ricinus-communis-seeds/isa.study.xlsx
 create mode 100644 studies/Ricinus-communis-seeds/protocols/.gitkeep
 create mode 100644 studies/Ricinus-communis-seeds/protocols/Plant-growth-conditions.md
 create mode 100644 studies/Ricinus-communis-seeds/resources/.gitkeep
 create mode 100644 workflows/.gitkeep

diff --git a/assays/2012_proteome_data_bmfz_stefanski/isa.assay.xlsx b/assays/2012_proteome_data_bmfz_stefanski/isa.assay.xlsx
index ff8e34df1423bc317e2f3be19ac62f7219c20603..0107750a77bcaaa4872f982921e8c815d27eec72 100644
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diff --git a/assays/2_2-FAME-analysis/README.md b/assays/2_2-FAME-analysis/README.md
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diff --git a/assays/2_2-FAME-analysis/protocols/.gitkeep b/assays/2_2-FAME-analysis/protocols/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/assays/2_2-FAME-analysis/protocols/2_2-FAME-analysis.md b/assays/2_2-FAME-analysis/protocols/2_2-FAME-analysis.md
new file mode 100644
index 0000000..7201a9c
--- /dev/null
+++ b/assays/2_2-FAME-analysis/protocols/2_2-FAME-analysis.md
@@ -0,0 +1,8 @@
+# 2.2 Fatty acid methylester (FAME) analysis
+
+The ricinoleic acid content in the castor bean endosperm was determined using gas chromatography-mass spectrometry (GC- MS)-based fatty acid methyl ester (FAME) method (Hielscher et al., 2017). Endosperm tissue was dissected from dry seeds, 24h- imbibed seeds and 1- to 6-day old dark-grown seedlings by removing the embryo and roots with the blunt end of a scalpel blade, weighed and shock frozen in liquid nitrogen.
+FAME analyses were generated by acid catalyzed methyl-ester formation using methanolic HCl as described by Hielscher et al. (2017). 20 mg endosperm was incubated at 90°C for one hour in 1 ml 3 N methanolic-HCl containing heptadecanoic acid (C17:0) as internal standard. The subsequent extraction of fatty acids from the sample was performed with 1 ml n-hexane and 1 ml 1% (w/v) sodium chloride. Samples were centrifuged at 2,000 g for five minutes. The resulting upper hexane phase was transferred into GC vials. The FAME extracts (1:100 diluted) were analyzed by GC Agilent 7890A gas chromatograph coupled to Waters GCT-TOF Premier mass spectrometer equipped with a Gerstel MPS2XL auto sampler.
+Quantification and identification of the detected fatty acids was carried out with QuantLynx and MassLynx, respectively. Peaks were integrated and the resulting areas used for determination of relative changes in the abundance of the castor oil-specific ricinoleic acid. As the Ricinus endosperm started to gain fresh weight during seedling development due to water uptake, the fatty acid quantities were normalized to the fresh weight of an average endosperm. All samples were analyzed in three biological replicates.
+
+
+- Hielscher, B., Charton, L., Mettler-Altmann, T., and Linka, N. (2017). Analysis of peroxisomal b-oxidation during storage oil mobilization in Arabidopsis thaliana Seedlings. Methods Mol. Biol. 1595, 291–304. doi: 10.1007/978-1-4939-6937-1_27
\ No newline at end of file
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diff --git a/studies/Ricinus-communis-seeds/protocols/.gitkeep b/studies/Ricinus-communis-seeds/protocols/.gitkeep
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diff --git a/studies/Ricinus-communis-seeds/protocols/Plant-growth-conditions.md b/studies/Ricinus-communis-seeds/protocols/Plant-growth-conditions.md
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+++ b/studies/Ricinus-communis-seeds/protocols/Plant-growth-conditions.md
@@ -0,0 +1,6 @@
+# 2.1 Plant growth conditions
+
+Dry seeds of Ricinus communis var. zanzibariensis were surface- sterilized in a solution of 0.1% (w/v) 8-quinolinol in water for ten minutes and soaked in running tap water over night for seed imbibition. The imbibed seeds were placed on moist vermiculite and incubated at 30°C in the dark (Beevers and Breidenbach, 1974).
+
+
+- Beevers, H., and Breidenbach, R. W. (1974). “Glyoxysomes,” in Methods in enzymology, Vol 16 Biomembranes, Part A. Eds. S. Fleciher and L. packer (New York London: Academic Press), 565–571.
\ No newline at end of file
diff --git a/studies/Ricinus-communis-seeds/resources/.gitkeep b/studies/Ricinus-communis-seeds/resources/.gitkeep
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diff --git a/workflows/.gitkeep b/workflows/.gitkeep
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-- 
GitLab