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Destiny is no matter of chance. It is a matter of choice.
It is not a thing to be waited for. It is a thing to be achieved.
William Jennings Bryan
Day 8: DNA design
Assignments Due
Overview of Experiment
Today you measure the fluorescence output of pTetR-GFP
mutants and compare to output of BBa_I13522.
YOU WILL HAVE TO
COME IN THE DAY BEFORE LAB DAY 8 AND PULL COLONIES:
BBa_I13522 (pTetR-GFP)
mutated RBS:
pick 5 colonies per team and grow overnight in 3
ml LB-Amp (50 µg/ml) at 37°C with shaking (250
rpm)
Experimental goal: How does mutation of the RBS affect
GFP output?
- Pellet cells as on lab day 7; remove LB and resuspend in
PBS; pellet a second time, remove PBS, and resuspend in PBS
- Plate undiluted cells cells in triplicate on a 96-well
plate
- Measure absorbance at 600 nm and measure fluorescence (excitation
at 501 nm / emission at 511 nm)
Copyright, Acknowledgements,
and Intended Use
Created by B. Beason (bbeason@rice.edu),
Rice University, 21 November 2007
Updated 13 November 2013