Team:Warsaw/RBSmeasurement

We have conducted measurement of various ribosome binding sites included in 2011 spring distribution. Our list of measured parts includes YFP E0030 , GFP SUPERFOLDER , RFP E1010, mOrange E2050, GFP E0040. DNA of SUPERFOLDER GFP, was included in the distribution as a part of a bigger biobrick. We obtained coding sequence of SUPERFOLDER GFP using PCR method. All measurements were conducted in psb1A2 - a high copy number vector that was used in previous RBS measurement experiments with GFP protein.

To measure fluorescence of samples 3 milliliters of LB with AMP were inoculated with a colony from the plate. Cultures were vigorously shaken in 37° C overnight. The next step was to centrifuge 1,5 milliliters of liquid culture in an 1,5 mltube. Pellet was resuspended in 100 microliters of RF. Finally, prepared samples were placed in the fluorimeter plate. We used Corning 96 deep well plates with clear UV transparent bottom. Measurements were carried out using Tecan HS2000 pro fluorimeter with bottom read mode and using active gain optimization function. Wavelength values for individual proteins: