Experiments

All the experiments we conducted

Wetlab


General Protocols


Agarose Gel Electrophoresis


SDS PAGE


Plasmid Isolation (commercial kit)


Making competent cells


Sequencing


Microvolume Spectrophotometer


Colony PCR


Gibson Assembly (commercial kit)


Transformation (E.coli)


Buffers and Media


All concentrations are given as final concentrations unless stated otherwise.

10x PBS


10x PBS


10x PBS


5x SDS loading buffer/dye


10x SDS running buffer


50x TAE buffer


HisTrap IMAC binding buffer


HisTrapIMAC elution buffer


HisTrapIMAC resuspension buffer


ITC buffer


LB/LB-agar (Miller) medium (1L)


M9 minimal medium (100 mL)


M9 salts 5x (1L)


LBD-PctD experiments

Expression of PctD-LBD


Cell lysis


Protein Purification


Dialysis


ITC


Chimeric receptor development


DNA fragments amplification (PCRs)


Plasmid assembly (Gibson assembly)


Transformation into E.coli strains (DH5a, VS1007)


Testing of Biosensor


Bacterial growth curves


Fluorescence microscopy


Quantification of Fluorescence in Plate Reader


Drylab


Binding Site Prediction with SSnet


SeeSar affinity prediction


Molecular Dynamic Simulation with GROMACS