From 0d65c6b0dc68c6e8aa75a5962ff88679c395d3a2 Mon Sep 17 00:00:00 2001
From: Viktoria Petrova <vipet103@hhu.de>
Date: Sat, 11 Jan 2025 16:09:12 +0100
Subject: [PATCH] add determination of bacterial titer assay and protocol

---
 .../DeterminationOfBacterialTiter/README.md   |   0
 .../dataset/.gitkeep                          |   0
 .../isa.assay.xlsx                            | Bin 0 -> 6907 bytes
 .../protocols/.gitkeep                        |   0
 .../DeterminationOfBacterialTiterProtocol.md  |   3 +++
 assays/MetaboliteAnalysis/isa.assay.xlsx      | Bin 16323 -> 16359 bytes
 6 files changed, 3 insertions(+)
 create mode 100644 assays/DeterminationOfBacterialTiter/README.md
 create mode 100644 assays/DeterminationOfBacterialTiter/dataset/.gitkeep
 create mode 100644 assays/DeterminationOfBacterialTiter/isa.assay.xlsx
 create mode 100644 assays/DeterminationOfBacterialTiter/protocols/.gitkeep
 create mode 100644 assays/DeterminationOfBacterialTiter/protocols/DeterminationOfBacterialTiterProtocol.md

diff --git a/assays/DeterminationOfBacterialTiter/README.md b/assays/DeterminationOfBacterialTiter/README.md
new file mode 100644
index 0000000..e69de29
diff --git a/assays/DeterminationOfBacterialTiter/dataset/.gitkeep b/assays/DeterminationOfBacterialTiter/dataset/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/assays/DeterminationOfBacterialTiter/isa.assay.xlsx b/assays/DeterminationOfBacterialTiter/isa.assay.xlsx
new file mode 100644
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diff --git a/assays/DeterminationOfBacterialTiter/protocols/.gitkeep b/assays/DeterminationOfBacterialTiter/protocols/.gitkeep
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diff --git a/assays/DeterminationOfBacterialTiter/protocols/DeterminationOfBacterialTiterProtocol.md b/assays/DeterminationOfBacterialTiter/protocols/DeterminationOfBacterialTiterProtocol.md
new file mode 100644
index 0000000..28f3725
--- /dev/null
+++ b/assays/DeterminationOfBacterialTiter/protocols/DeterminationOfBacterialTiterProtocol.md
@@ -0,0 +1,3 @@
+## Determination of bacterial titer
+
+Bacterial titer in the roots was determined using the qPCR method as described in Koprivova et al. (2023). Genomic DNA was extracted by a buffer containing 0.025 M EDTA, 0.2 M Tris pH 8.0, 0.25 M NaCl, and 0.5% SDS. After 10 min incubation at 65 °C and centrifugation, the supernatant was precipitated with isopropanol, washed with 70% ethanol, and resuspended in 100 µl of sterile water. For the qPCR, 13 ng of corresponding DNA samples were used with *Arabidopsis*- (At primer AT4G26410) and *B. glumae* PG1-specific primer (Burk1 for NR042931). The qPCR conditions were the same as for expression analysis. The qPCRs were performed in duplicate for each of the four independent samples. The qPCR results were related to the bacterial titer as established in Koprivova et al. (2023).
\ No newline at end of file
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