From 95da09291b507f7eb0a5e17c3861c3c8dd0c525a Mon Sep 17 00:00:00 2001
From: Gesa Helmsorig <gesa.helmsorig@hhu.de>
Date: Wed, 30 Oct 2024 16:29:21 +0100
Subject: [PATCH] added metadata

---
 assays/DNA_extraction/isa.assay.xlsx          | Bin 25344 -> 23845 bytes
 assays/MSA_dissection/isa.assay.xlsx          | Bin 23241 -> 23241 bytes
 assays/biparental_mapping/isa.assay.xlsx      | Bin 46857 -> 46856 bytes
 assays/cDNA_synthesis/isa.assay.xlsx          | Bin 27342 -> 27341 bytes
 assays/embryo_transformation/isa.assay.xlsx   | Bin 13664 -> 11337 bytes
 assays/leaf_sampling/isa.assay.xlsx           | Bin 44376 -> 44375 bytes
 .../dataset/T0_genotype_results.xlsx          | Bin 6602 -> 9896 bytes
 .../dataset/T1_genotype_results.xlsx          | Bin 6602 -> 10556 bytes
 assays/mutant_genotyping/isa.assay.xlsx       | Bin 11258 -> 14815 bytes
 .../protocols/plant_genotyping.md             |  11 +++++++++++
 .../protocols/plant_genotyping.txt            |   4 ----
 assays/plant_phenotyping/isa.assay.xlsx       | Bin 38558 -> 32487 bytes
 assays/plant_propagation/isa.assay.xlsx       | Bin 10981 -> 13025 bytes
 .../protocols/progagation_of_mutant_plants.md |   0
 studies/MSA_dissection_eam7/isa.study.xlsx    | Bin 24075 -> 24075 bytes
 .../isa.study.xlsx                            | Bin 21674 -> 21673 bytes
 .../isa.study.xlsx                            | Bin 9916 -> 17109 bytes
 .../generating_lwd1_mutants/isa.study.xlsx    | Bin 13259 -> 11117 bytes
 .../genotyping_lwd1_mutants/isa.study.xlsx    | Bin 7509 -> 10830 bytes
 19 files changed, 11 insertions(+), 4 deletions(-)
 create mode 100644 assays/mutant_genotyping/protocols/plant_genotyping.md
 delete mode 100644 assays/mutant_genotyping/protocols/plant_genotyping.txt
 create mode 100644 assays/plant_propagation/protocols/progagation_of_mutant_plants.md

diff --git a/assays/DNA_extraction/isa.assay.xlsx b/assays/DNA_extraction/isa.assay.xlsx
index 6a4e6d819baefd2c48387bac8e40a8acc99c14ab..0cecddb6a347a5271246edfcad4b379dd30eabf6 100644
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diff --git a/assays/MSA_dissection/isa.assay.xlsx b/assays/MSA_dissection/isa.assay.xlsx
index 4ba043f0c5b7713af801b3bcbf62691dfaaa80b6..43ce3d91b80895b18480a43f06ac9e89f6782248 100644
GIT binary patch
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diff --git a/assays/biparental_mapping/isa.assay.xlsx b/assays/biparental_mapping/isa.assay.xlsx
index 86cf18bbaec63ee37127adc445443140a0afe41f..398e732c136a2e96195371dd60fe07144f344421 100644
GIT binary patch
delta 772
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diff --git a/assays/cDNA_synthesis/isa.assay.xlsx b/assays/cDNA_synthesis/isa.assay.xlsx
index 897d0e9596ad7ef56e9bedf03221b47b057de06c..ae7c6c0a977bc582f34929a16ae5b03f38fa068b 100644
GIT binary patch
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diff --git a/assays/embryo_transformation/isa.assay.xlsx b/assays/embryo_transformation/isa.assay.xlsx
index e37966eb7f619d604aab251bda64a6390cb12a63..6ff98f841fb28c944b0a5afbe4de9040540942fb 100644
GIT binary patch
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diff --git a/assays/mutant_genotyping/dataset/T1_genotype_results.xlsx b/assays/mutant_genotyping/dataset/T1_genotype_results.xlsx
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diff --git a/assays/mutant_genotyping/isa.assay.xlsx b/assays/mutant_genotyping/isa.assay.xlsx
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diff --git a/assays/mutant_genotyping/protocols/plant_genotyping.md b/assays/mutant_genotyping/protocols/plant_genotyping.md
new file mode 100644
index 0000000..0a4205e
--- /dev/null
+++ b/assays/mutant_genotyping/protocols/plant_genotyping.md
@@ -0,0 +1,11 @@
+**lwd1 T0 and T1 plants **
+
+- DNA was extracted from leaves of all mutants using QIAGEN_DNeasy-Plant_mini protocol
+- plants were tested for possible mutations in the CDS of LWD1 by amplifying the full genomic sequence using the primers LWD1_71us_F (5'-ATCAGACCCCTCCAACGACT-3') and LWD1_21ds_R (5'-TTCCTCTTTCTACCCCACCA-3')
+- the PCR fragments were Sanger sequenced using different primers:
+	- plants transformed with pMGE599/7-97 (approach 1): LWD1_71us_F 
+	- plants transformed with pMGE599/182-1222 (approach 2): LWD1_qPCR114F (5'-CAGCTCATAGCCCATGACAA-3')
+- mutations and the zygosity of the mutations were determined relative to the start codon of LWD1
+
+**only in T1 plants**:
+- PCR was performed on transformation vector to check presence/absence of Cas9 using the primers pMGE599_M13F (5'-GTTTTCCCAGTCACGAC-3') and pMGE599_R (5'-TGTCGACATCGATAAGCTTGAA-3') 
\ No newline at end of file
diff --git a/assays/mutant_genotyping/protocols/plant_genotyping.txt b/assays/mutant_genotyping/protocols/plant_genotyping.txt
deleted file mode 100644
index 6354cb6..0000000
--- a/assays/mutant_genotyping/protocols/plant_genotyping.txt
+++ /dev/null
@@ -1,4 +0,0 @@
-- DNA extracted from plants using QIAGEN protocol
-- PCR done with primers lwd f/r
-- sent for Sanger sequencing
-- Sequences were evaluated: mutations in CDS of lwd1?
\ No newline at end of file
diff --git a/assays/plant_phenotyping/isa.assay.xlsx b/assays/plant_phenotyping/isa.assay.xlsx
index 3c09b64b3e0918cc5b1919e5af250d695ff43abe..529fd1b3db586328e02efdd863a4af9b81f98c3a 100644
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diff --git a/assays/plant_propagation/isa.assay.xlsx b/assays/plant_propagation/isa.assay.xlsx
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diff --git a/assays/plant_propagation/protocols/progagation_of_mutant_plants.md b/assays/plant_propagation/protocols/progagation_of_mutant_plants.md
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diff --git a/studies/gene_expression_lwd1_mutants_diurnal/isa.study.xlsx b/studies/gene_expression_lwd1_mutants_diurnal/isa.study.xlsx
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diff --git a/studies/generating_lwd1_mutants/isa.study.xlsx b/studies/generating_lwd1_mutants/isa.study.xlsx
index 17e39f26a40c929bef6f6acd9d86966e7d461d8a..02969f3fafa6a6c1cbd1eeb5b8f6e5813113d234 100644
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diff --git a/studies/genotyping_lwd1_mutants/isa.study.xlsx b/studies/genotyping_lwd1_mutants/isa.study.xlsx
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