diff --git a/assays/BGC prediction using antiSMASH/isa.assay.xlsx b/assays/BGC prediction using antiSMASH/isa.assay.xlsx index ab438f400679e35e0de49967e3a6af6f9a499bb5..b06b8354d92b038d824c16bc5ed5745d5632e109 100644 Binary files a/assays/BGC prediction using antiSMASH/isa.assay.xlsx and b/assays/BGC prediction using antiSMASH/isa.assay.xlsx differ diff --git a/assays/DetectionOfR401DAPGandPyoverdine/isa.assay.xlsx b/assays/DetectionOfR401DAPGandPyoverdine/isa.assay.xlsx index 69890d3be81d994fb8a52d2a22db9d79c3b0b62c..e846156dd260e8768b142f4e0c7d8c94e6b7ff3c 100644 Binary files a/assays/DetectionOfR401DAPGandPyoverdine/isa.assay.xlsx and b/assays/DetectionOfR401DAPGandPyoverdine/isa.assay.xlsx differ diff --git a/assays/Metabolomics Analysis/isa.assay.xlsx b/assays/Metabolomics Analysis/isa.assay.xlsx index 3419fe3fdb9b0a78280c93f59985e6a6327c7c18..c876c4ee3472462539cb9506cce354ad57473dfa 100644 Binary files a/assays/Metabolomics Analysis/isa.assay.xlsx and b/assays/Metabolomics Analysis/isa.assay.xlsx differ diff --git a/assays/Mutant Generation/isa.assay.xlsx b/assays/Mutant Generation/isa.assay.xlsx index dd96ac643f307d69ab419072c2fa5e8111b16456..0d4d9bfb148129fe76665a291dfab96bad264f67 100644 Binary files a/assays/Mutant Generation/isa.assay.xlsx and b/assays/Mutant Generation/isa.assay.xlsx differ diff --git a/assays/Mutant Generation/protocols/TransformationBW29427.md b/assays/Mutant Generation/protocols/TransformationBW29427.md new file mode 100644 index 0000000000000000000000000000000000000000..232c0db9686d78004af83fb2537e2dfa74d167b6 --- /dev/null +++ b/assays/Mutant Generation/protocols/TransformationBW29427.md @@ -0,0 +1,3 @@ +## Transformation into chemically competent E. coli BW29427 cells + +The vector was transformed into 50 μl chemically competent BW29427 *E. coli* cells according to the following heat shock protocol: 2 μl of the vector were gently mixed with 50 μl of competent cells, and the resulting mixture was incubated on ice for 30 min. The mixture was transferred to a water bath at 42 °C for 1 min and put back on ice for 2 min. Then, 1 ml of 50% TSB with 50 μg/ml diaminopimelic acid (DAP; Sigma-Aldrich) was added to the heat-shocked cells, the mixture was left to regenerate at 37 °C for 1 h and then plated on 50% TSA containing 25 μg/ml Kanamycin (Kan) and 50 μg/ml DAP. The plates were incubated at 37 °C overnight. Resulting colonies were validated by colony PCR using the M13F and M13R primers. Colony PCR was performed on at least four separate colonies with 0.4 μl DFS-Taq polymerase (BIORON) in 25 μl reactions containing 2.5 μl 10x incomplete buffer (BIORON), 0.5 10 mM MgC12, 0.5 μl 10 mM dNTPs, 0.75 μl 10 μM forward primer, 0.75 μl 10 μM reverse primer, a small fraction of a colony and filled up to 25 μl with nuclease-free water. The tubes were placed in a thermocycler set at the following program: 94°C for 2 min, 35 cycles of 94°C for 30 s, 55°C for 30 s, 72°C for 2 min, then a final extension at 72 °C for 10 min. Five microliters of the PCR product were combined with 1 μl Orange DNA Loading Dye and analysed by DNA agarose electrophoresis. Positive colonies were purified by streaking on new 50% TSA plates containing 25 μg/ml Kan and 50 μg/ml DAP and further verified by Sanger sequencing (Eurofins Scientific) following the manufacturer's protocol. diff --git a/assays/Screen for antagonistic interbacterial interactions/isa.assay.xlsx b/assays/Screen for antagonistic interbacterial interactions/isa.assay.xlsx index 5c996e6d84d020f9c80090083ac9f00e1d7d1763..c5fbcc9dda8928f7c4f7b86087023549c3be61da 100644 Binary files a/assays/Screen for antagonistic interbacterial interactions/isa.assay.xlsx and b/assays/Screen for antagonistic interbacterial interactions/isa.assay.xlsx differ diff --git a/isa.investigation.xlsx b/isa.investigation.xlsx index ba6e39478168c92e5be36ba5756b6f5079d10710..b15f1a4b4e624c8fb1fa381bd79da504ac15d5de 100644 Binary files a/isa.investigation.xlsx and b/isa.investigation.xlsx differ diff --git a/studies/Bacterial culture conditions/isa.study.xlsx b/studies/Bacterial culture conditions/isa.study.xlsx index e6aa52d765bb890d581bcbb4eb1d33dd61ecada6..f79bce8581a85dd69170620fa7fb175e0bed438c 100644 Binary files a/studies/Bacterial culture conditions/isa.study.xlsx and b/studies/Bacterial culture conditions/isa.study.xlsx differ diff --git a/studies/Plant Growth Conditions/isa.study.xlsx b/studies/Plant Growth Conditions/isa.study.xlsx index 293a48d741bfac0803a331bcaa4f1e559d6bf046..36f4e75c4a78fa0fdeaa5b15460c17377974b9a2 100644 Binary files a/studies/Plant Growth Conditions/isa.study.xlsx and b/studies/Plant Growth Conditions/isa.study.xlsx differ