From 3e7b99cbede6d01ee5ab9c8374edcb0f0cdfad7c Mon Sep 17 00:00:00 2001
From: Viktoria Petrova <vipet103@hhu.de>
Date: Mon, 8 Jan 2024 19:14:17 +0100
Subject: [PATCH] add qRT-PCR protocol

---
 assays/qRT-PCR/protocols/qRT-PCR.md                             | 2 ++
 ...endipitaIndicaSerendipitaVermiferaAndBipolarisSorokiniana.md | 2 +-
 2 files changed, 3 insertions(+), 1 deletion(-)
 create mode 100644 assays/qRT-PCR/protocols/qRT-PCR.md

diff --git a/assays/qRT-PCR/protocols/qRT-PCR.md b/assays/qRT-PCR/protocols/qRT-PCR.md
new file mode 100644
index 0000000..f6147b3
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+++ b/assays/qRT-PCR/protocols/qRT-PCR.md
@@ -0,0 +1,2 @@
+## qRT-PCR
+To quantify fungal colonization and *PR10* gene expression, RNA extraction, cDNA generation and RT-PCR were performed as previously described (Sarkar et al., 2019).
\ No newline at end of file
diff --git a/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/protocols/SerendipitaIndicaSerendipitaVermiferaAndBipolarisSorokiniana.md b/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/protocols/SerendipitaIndicaSerendipitaVermiferaAndBipolarisSorokiniana.md
index da1b0f2..56f170b 100644
--- a/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/protocols/SerendipitaIndicaSerendipitaVermiferaAndBipolarisSorokiniana.md
+++ b/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/protocols/SerendipitaIndicaSerendipitaVermiferaAndBipolarisSorokiniana.md
@@ -1,2 +1,2 @@
 ## Serendipita indica, Serendipita vermifera and Bipolaris sorokiniana
-The growth conditions for barley, *S. indica* (DSM11827, Si), *S. vermifera* (MAFF305830, Sv) and *B. sorokiniana* (ND90Pr, Bs) and the preparation of fungal suspensions for plant inoculation have been described previously.41,75 Four-day-old barley seedlings were transferred to sterile jars on 1/10 PNM (pH 5.7) and inoculated with 3 mL of either sterile water as control, *Si* chlamydospores (500,000 spores·mL-1), *Sv* mycelium (1 g per 50 mL), or *Bs* conidia (5,000 spores·mL-1). Plants were grown on a day/night cycle of 16/8 h at 22/18 °C and 60% humidity under a light intensity of 108 μmol·m−2·s−1. Plant roots were harvested six days post inoculation (dpi), washed thoroughly to remove extraradical fungal hyphae, and frozen in liquid nitrogen. Four barley plants were pooled per biological replicate. To quantify fungal colonization and *PR10* gene expression, RNA extraction, cDNA generation and RT-PCR were performed as previously described.41
\ No newline at end of file
+The growth conditions for barley, *S. indica* (DSM11827, Si), *S. vermifera* (MAFF305830, Sv) and *B. sorokiniana* (ND90Pr, Bs) and the preparation of fungal suspensions for plant inoculation have been described previously.41,75 Four-day-old barley seedlings were transferred to sterile jars on 1/10 PNM (pH 5.7) and inoculated with 3 mL of either sterile water as control, *Si* chlamydospores (500,000 spores·mL-1), *Sv* mycelium (1 g per 50 mL), or *Bs* conidia (5,000 spores·mL-1). Plants were grown on a day/night cycle of 16/8 h at 22/18 °C and 60% humidity under a light intensity of 108 μmol·m−2·s−1. Plant roots were harvested six days post inoculation (dpi), washed thoroughly to remove extraradical fungal hyphae, and frozen in liquid nitrogen. Four barley plants were pooled per biological replicate. 
\ No newline at end of file
-- 
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