From f3fdd37b8017e6c643dd91dfb08c3688fedac343 Mon Sep 17 00:00:00 2001
From: Sabrina Zander <sabrina.zander@uni-duesseldorf.de>
Date: Tue, 21 Jan 2025 11:23:37 +0100
Subject: [PATCH] add assay 1-2

---
 assays/1-2_qPCR-taggedVariants/README.md       |   1 +
 .../1-2_qPCR-taggedVariants/dataset/.gitkeep   |   0
 .../20210304_qPCR_DtxR_Tag_Evaluation.xlsx     | Bin 0 -> 19504 bytes
 .../dataset/20210403_qPCR_DtxR_Tags.rtp        | Bin 0 -> 891328 bytes
 .../dataset/qPCR_DtxRtags.png                  | Bin 0 -> 31558 bytes
 assays/1-2_qPCR-taggedVariants/isa.assay.xlsx  | Bin 0 -> 11823 bytes
 .../1-2_qPCR-taggedVariants/protocols/.gitkeep |   0
 .../protocols/qPCR_method.txt                  |   1 +
 8 files changed, 2 insertions(+)
 create mode 100644 assays/1-2_qPCR-taggedVariants/README.md
 create mode 100644 assays/1-2_qPCR-taggedVariants/dataset/.gitkeep
 create mode 100644 assays/1-2_qPCR-taggedVariants/dataset/20210304_qPCR_DtxR_Tag_Evaluation.xlsx
 create mode 100644 assays/1-2_qPCR-taggedVariants/dataset/20210403_qPCR_DtxR_Tags.rtp
 create mode 100644 assays/1-2_qPCR-taggedVariants/dataset/qPCR_DtxRtags.png
 create mode 100644 assays/1-2_qPCR-taggedVariants/isa.assay.xlsx
 create mode 100644 assays/1-2_qPCR-taggedVariants/protocols/.gitkeep
 create mode 100644 assays/1-2_qPCR-taggedVariants/protocols/qPCR_method.txt

diff --git a/assays/1-2_qPCR-taggedVariants/README.md b/assays/1-2_qPCR-taggedVariants/README.md
new file mode 100644
index 0000000..f28daf6
--- /dev/null
+++ b/assays/1-2_qPCR-taggedVariants/README.md
@@ -0,0 +1 @@
+Within the publication, the qPCR only for the ::dtxR-His-Cterm-2link variant is depicted, as the Strep version was not used due to disturbances.
\ No newline at end of file
diff --git a/assays/1-2_qPCR-taggedVariants/dataset/.gitkeep b/assays/1-2_qPCR-taggedVariants/dataset/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/assays/1-2_qPCR-taggedVariants/dataset/20210304_qPCR_DtxR_Tag_Evaluation.xlsx b/assays/1-2_qPCR-taggedVariants/dataset/20210304_qPCR_DtxR_Tag_Evaluation.xlsx
new file mode 100644
index 0000000000000000000000000000000000000000..2645f0138fe178235db32523413df3b23744eb1a
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diff --git a/assays/1-2_qPCR-taggedVariants/dataset/20210403_qPCR_DtxR_Tags.rtp b/assays/1-2_qPCR-taggedVariants/dataset/20210403_qPCR_DtxR_Tags.rtp
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diff --git a/assays/1-2_qPCR-taggedVariants/protocols/.gitkeep b/assays/1-2_qPCR-taggedVariants/protocols/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/assays/1-2_qPCR-taggedVariants/protocols/qPCR_method.txt b/assays/1-2_qPCR-taggedVariants/protocols/qPCR_method.txt
new file mode 100644
index 0000000..4cbd779
--- /dev/null
+++ b/assays/1-2_qPCR-taggedVariants/protocols/qPCR_method.txt
@@ -0,0 +1 @@
+For analysis of the C. glutamicum strain with a tagged DtxR variant (::dtxR-C-linker-His), cultivation in deep-well plates (standard, 36 µM FeSO4) or flasks (iron excess, 100 µM FeSO4) was performed, harvesting cells in ice-falcons at an OD600 of ~5 in exponential phase. Using the Luna One-Step RT-qPCR Kit (New England BioLabs, Frankfurt am Main) according to manufacturer’s instructions, qPCR was performed in the qTower (Analytik Jena, Jena). As a reference gene for normalization, the housekeeping gene ddh was used, besides the target gene dtxR. Analysis followed using qPCRsoft 3.1 (Analytik Jena, Jena) and fold-change was calculated according to the 2-ΔΔCt method.
\ No newline at end of file
-- 
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