Team:William and Mary/Experiments
Methods & Data
Overview
In the interest of promoting reproducible science, and enabling others to build off of our work, we have included both the precise methods used as well as the raw data (in csv format) for every experiment mentioned on our wiki. Experiments are split up by the page on which they are mentioned. For experiments performed with flow cytometry, .fcs files for both measurements and calibration beads (Spherotek RCP-30-5A, lot number: AB01) are included as a separate file. Whenever used, the term biological replicates refers to a distinct colony from a transformation of the same miniprep.
For all experiments, unless stated otherwise, the following is true: Circuits used were present in psb1C3, antibiotics were used in the following concentrations [Chloramphenicol: 7.5mg/mL, Kanamycin: 35mg/mL], the media used was M9 minimal media with 0.4% Casamino Acids and 0.4% Glucose (see protocols), 96 well plates used with the microplate reader (Synergy H1 [Biotek]) were Black Griener Fluotrack 200 flat bottom plates with the matching lid, the microplate reader was set to linear shaking at max speed and gradient temperatures were used to prevent condensation.
Heat Inducible System
Figure 2: In both cases colonies were picked into 200µL of media into a 96 well plate and cultures were grown at 29C until cultures showed OD600s consistent with mid-log growth. Cultures were then diluted to an OD600 of ~.05 and then subjected to their conditions. For A, this was a 37C for the entire period, whereas for B this was 37C from 160 minutes to 280 minutes and 29C elsewhere. Data
Figure 2: In both cases colonies were picked into 200µL of media into a 96 well plate and cultures were grown at 29C until cultures showed OD700s consistent with mid-log growth. Cultures were then diluted to an OD600 of ~.05 and then subjected to their conditions. For A, this was a 37C for the entire period, whereas for B this was 37C from 140 minutes to 280 minutes and 29C elsewhere. Data