Cell measurement protocol

1. Transform E. coli DH5α with these following plasmids:

     a) Positive Control

     b) Negative Control

     c) Test Device 1

     d) Test Device 2

     e) Test Device 3

     f) Test Device 4

     g) Test Device 5

     h) Test Device 6

2. Pick 2 colonies from each plate and inoculate it on 5 mL LB medium + Chloramphenicol. Grow the cells overnight (16 hours) at 37°C and 220 rpm.

3. Cell growth, sampling, and assay

     a) Set your instrument to read OD600 (as OD calibration setting)

     b) Measure OD600 of the overnight cultures

     c) Record data

     d) Import data into Excel (normalisation tab) Sheet_1 provided

     e) Dilute the cultures to a target OD600 of 0.02 in 10 ml LB medium + Chloramphenicol in 50 mL falcon tube

     f) Take 100μL (1% of total volume) samples of the cultures in the 96-well plates, Use Thermo VARIOSKAN FLASH to shake the 96-well plates, measure the samples (OD and FlTC) at 0, 1, 2, 3, 4, 5, and 6 hours of incubation

     g) Record data in the notebook

     h) Import data into Excel (cell measurement tab) Sheet_1 provided