Protocols

Cell culturing protocols

CC1: Miniprep
CC2: Midiprep
CC3: Making cells competent
CC4: Ligation
CC5: Transformation into BL21 using electrical shock
CC6: Transformation into Top10 using chemical competence
CC7: Making LB plates
CC8: Extracting DNA from the registry
CC9: Preparation of XL1-Blue electrocompetent cells & XL1-Blue quality control
CC10: PCR
CC11: SLIC
CC12: Biobricking parts from oligios
CC13: Genome Prep for Restriction Assay

Calibrations

CA1: Optical Density Calibration
GFP fluorescence calibration
- CA2: External GFP fluorescence
- CA3: Intracellular GFP fluorescence

Autoinduction

AI1: Secondary Carbon Source Experiment
AI2: Glucose Time Delay

Protein Production

PP1: IPTG Toxicity
PP2: IPTG characterisation
PP3: Cellulase
PP4: PAH

Encapsulation

EN1: Colanic acid production amount
EN2: Colanic acid protection from pH
[[Team:Imperial_College_London/Wetlab/Protocols/Trehalose | EN3: Trehalose production

Killing

KI1: Thermoinduction
KI2: Restriction Enzyme activity

Autoinduction assays

Lac characterisation and IPTG effect on protein production

Aims

Characterise Lac promoter by varying amounts of IPTG
Determine the IPTG concentration that allows for maximal protein production while still being non-toxic to the cell

Assay

The cells will be grown until OD=0.7. Now, IPTG of various concentrations will be added, and the RFP output will be measured.
The experiment will generate OD and fluoresence data for RFP
The secondary carbon source will be taken from the previous experiment (see Secondary carbon source selection)
The glucose concentration will be taken from commercial autoinduction media (0.05%) – takes about 7 hours to exhaust

See the protocol for more details