diff --git a/assays/1-2_qPCR-taggedVariants/README.md b/assays/1-2_qPCR-taggedVariants/README.md new file mode 100644 index 0000000000000000000000000000000000000000..f28daf6f7a0baea74754b9f164aa9aea820c0c82 --- /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 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 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 Binary files /dev/null and b/assays/1-2_qPCR-taggedVariants/dataset/20210304_qPCR_DtxR_Tag_Evaluation.xlsx differ 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 new file mode 100644 index 0000000000000000000000000000000000000000..ea4c783414a90eaef8927b136adf8ee5db2eaa03 Binary files /dev/null and b/assays/1-2_qPCR-taggedVariants/dataset/20210403_qPCR_DtxR_Tags.rtp differ diff --git a/assays/1-2_qPCR-taggedVariants/dataset/qPCR_DtxRtags.png b/assays/1-2_qPCR-taggedVariants/dataset/qPCR_DtxRtags.png new file mode 100644 index 0000000000000000000000000000000000000000..335854b047b5a02a9c6dbe9fd2ac7bd836e6b1db Binary files /dev/null and b/assays/1-2_qPCR-taggedVariants/dataset/qPCR_DtxRtags.png differ diff --git a/assays/1-2_qPCR-taggedVariants/isa.assay.xlsx b/assays/1-2_qPCR-taggedVariants/isa.assay.xlsx new file mode 100644 index 0000000000000000000000000000000000000000..6c8da90ace6455f848e406dce273ab3c98b600c3 Binary files /dev/null and b/assays/1-2_qPCR-taggedVariants/isa.assay.xlsx differ diff --git a/assays/1-2_qPCR-taggedVariants/protocols/.gitkeep b/assays/1-2_qPCR-taggedVariants/protocols/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 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 0000000000000000000000000000000000000000..4cbd779d45d513e483e76eb9707276d538a7d606 --- /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