From f140421524dc3d5a95b553b0ad7605fbd49d450b Mon Sep 17 00:00:00 2001
From: Viktoria Petrova <vipet103@hhu.de>
Date: Mon, 25 Dec 2023 13:58:44 +0100
Subject: [PATCH] add EnzymaticCarbohydrateDigestionAndTLC assay

---
 .../README.md                                 |   0
 .../dataset/.gitkeep                          |   0
 .../isa.assay.xlsx                            | Bin 0 -> 6915 bytes
 .../protocols/.gitkeep                        |   0
 .../EnzymaticCarbohydrateDigestion.md         |   2 ++
 .../protocols/TLC.md                          |   2 ++
 .../isa.assay.xlsx                            | Bin 7020 -> 7021 bytes
 assays/OxidativeBurstAssay/isa.assay.xlsx     | Bin 6899 -> 6900 bytes
 isa.investigation.xlsx                        | Bin 11044 -> 11077 bytes
 studies/Barley/isa.study.xlsx                 | Bin 7676 -> 7717 bytes
 .../isa.study.xlsx                            | Bin 7535 -> 7536 bytes
 .../isa.study.xlsx                            | Bin 7517 -> 7518 bytes
 12 files changed, 4 insertions(+)
 create mode 100644 assays/EnzymaticCarbohydrateDigestionAndTLC/README.md
 create mode 100644 assays/EnzymaticCarbohydrateDigestionAndTLC/dataset/.gitkeep
 create mode 100644 assays/EnzymaticCarbohydrateDigestionAndTLC/isa.assay.xlsx
 create mode 100644 assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/.gitkeep
 create mode 100644 assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/EnzymaticCarbohydrateDigestion.md
 create mode 100644 assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md

diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/README.md b/assays/EnzymaticCarbohydrateDigestionAndTLC/README.md
new file mode 100644
index 0000000..e69de29
diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/dataset/.gitkeep b/assays/EnzymaticCarbohydrateDigestionAndTLC/dataset/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/isa.assay.xlsx b/assays/EnzymaticCarbohydrateDigestionAndTLC/isa.assay.xlsx
new file mode 100644
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diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/.gitkeep b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/EnzymaticCarbohydrateDigestion.md b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/EnzymaticCarbohydrateDigestion.md
new file mode 100644
index 0000000..7b4ecbf
--- /dev/null
+++ b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/EnzymaticCarbohydrateDigestion.md
@@ -0,0 +1,2 @@
+## Enzymatic carbohydrate digestion
+Carbohydrate digestion assays were performed using either purified barley GBP (heterologously expressed in *N. benthamiana*) or barley BGLUII33 (available from Megazyme, E-LAMHV). Preparations of the fungal CW and EPS matrix were incubated overnight in sterile Milli-Q water at 65 °C prior to enzymatic digestion. Substrate and enzyme concentrations, buffer compositions, digestion temperature and time are described in the figure captions. Digestion was stopped by denaturing the enzymes at 95 °C for 10 min and the digestion products were stored at -20 °C prior to use.
\ No newline at end of file
diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md
new file mode 100644
index 0000000..a700062
--- /dev/null
+++ b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md
@@ -0,0 +1,2 @@
+## Thin layer chromatography (TLC)
+An aliquot of each sample was subjected to TLC using a silica gel 60 F254 aluminum TLC plate (Merck Millipore, Burlington, USA), using a running buffer containing ethyl acetate/acetic acid/methanol/formic acid/water at a ratio of 8:4:1:1:1 (v/v). D-glucose, laminaribiose β-1-3-(Glc)2, laminaritriose β-1-3-(Glc)3, gentiobiose β-1-6-(Glc)2, and laminaripentaose β-1-3-(Glc)5 at a concentration of 1.5 mg·mL−1 were used as standards (Megazyme, Bray, Ireland). To visualize the glucan fragments, the TLC plate was sprayed with glucan developer solution (45 mg N-naphthol, 4.8 mL H2SO4, 37.2 mL ethanol and 3 mL water) and baked at 95 °C until the glucan bands became visible (approximately 4-5 min).
\ No newline at end of file
diff --git a/assays/MS-MSAnalysisOfThePull-downProteins/isa.assay.xlsx b/assays/MS-MSAnalysisOfThePull-downProteins/isa.assay.xlsx
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diff --git a/assays/OxidativeBurstAssay/isa.assay.xlsx b/assays/OxidativeBurstAssay/isa.assay.xlsx
index 079eac191b001fc238d086b7f8a5b0bdfb056aa3..d541a79ab966c00422257b31faf24c6420cd1f69 100644
GIT binary patch
delta 654
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diff --git a/isa.investigation.xlsx b/isa.investigation.xlsx
index 1c5b25406e0b0196214bd30d95f23270918796e0..c182d3e5edbd8c67554fa7b660b50bb64ce0dfce 100644
GIT binary patch
delta 5308
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diff --git a/studies/Barley/isa.study.xlsx b/studies/Barley/isa.study.xlsx
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GIT binary patch
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diff --git a/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/isa.study.xlsx b/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/isa.study.xlsx
index 9746ba5b1312c8b45bb946138b741920ac29cf1e..c6c19e65eb862ae646b4a80b7b296fb77b3b7186 100644
GIT binary patch
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diff --git a/studies/PlantMaterialAndGrowthConditions/isa.study.xlsx b/studies/PlantMaterialAndGrowthConditions/isa.study.xlsx
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-- 
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