From 5a3472f5273121a17787716c4e0c2b1355a2f7ed Mon Sep 17 00:00:00 2001
From: Viktoria Petrova <vipet103@hhu.de>
Date: Sat, 15 Jun 2024 12:00:35 +0200
Subject: [PATCH] add protocol reference to isa study #13

---
 .../protocols/PlantMaterialAndGrowthConditions.md     | 11 ++++++++++-
 1 file changed, 10 insertions(+), 1 deletion(-)

diff --git a/studies/HordeumVulgare/protocols/PlantMaterialAndGrowthConditions.md b/studies/HordeumVulgare/protocols/PlantMaterialAndGrowthConditions.md
index 27895a9..42dc1cb 100644
--- a/studies/HordeumVulgare/protocols/PlantMaterialAndGrowthConditions.md
+++ b/studies/HordeumVulgare/protocols/PlantMaterialAndGrowthConditions.md
@@ -1,3 +1,12 @@
 ## PlantMaterialAndGrowthConditions
 
-All experiments, including the generation of CRISPR/Cas9 knock-out lines, were performed with the spring barley (*H. vulgare L.*) cv. Golden Promise Fast, an introgression line carrying the Ppd-H1 allele that confers fast flowering (Gol et al., 2021) From here on, we use “control” to name this non-mutagenized cultivar that carries normal copies of *GBP1* and *GBP2*. For ROS burst assays, barley seeds were surface sterilized with 6% sodium hypochlorite for 1 h and then washed extensively (5 × 30 mL sterile water). Seeds were germinated on wet filter paper at room temperature in the dark under sterile conditions for three days before transfer to sterile jars containing solid 1/10 plant nutrition medium (PNM), pH 5.7 and 0.4% Gelrite (Duchefa, Haarlem, the Netherlands). Seedlings were cultured for four days in a growth chamber under long-day conditions (day/night cycle of 16/8 h, 22 °C/18 °C, light intensity of 108 μmol·m−2·s−1).
\ No newline at end of file
+All experiments, including the generation of CRISPR/Cas9 knock-out lines, were performed with the spring barley (*H. vulgare L.*) cv. Golden Promise Fast, an introgression line carrying the Ppd-H1 allele that confers fast flowering (Gol et al., 2021) From here on, we use “control” to name this non-mutagenized cultivar that carries normal copies of *GBP1* and *GBP2*. 
+
+ROS burst assay
+For ROS burst assays, barley seeds were surface sterilized with 6% sodium hypochlorite for 1 h and then washed extensively (5 × 30 mL sterile water). 
+
+Germination
+Seeds were germinated on wet filter paper at room temperature in the dark under sterile conditions for three days before transfer to sterile jars containing solid 1/10 plant nutrition medium (PNM), pH 5.7 and 0.4% Gelrite (Duchefa, Haarlem, the Netherlands). 
+
+Growth chamber
+Seedlings were cultured for four days in a growth chamber under long-day conditions (day/night cycle of 16/8 h, 22 °C/18 °C, light intensity of 108 μmol·m−2·s−1).
\ No newline at end of file
-- 
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