Team:TU Delft/Project/rbs-characterization/characterization

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Characterization

The cells were cultured over 18 hours in 96-well plates using a Gen5 fluorescence and absorbance plate reader, LB with ampicillin was the culture medium used in these experiments. Only the results obtained during exponential growth phase were taken into account.

26 07 2010 Rbs.png

Note: Fluorescence (y-axis) is reported as Arbitrary Fluorescence Units.

The RBS strength was calculated by taking the mean of the ratio between the expression of the standard RBS (B0032) and expression of Anderson RBS over time. The Expression is defined as the quotient of the measured fluorescence divided by measured biomass (OD at 600nm).


RBS Strength
J61100 0.047513
J61101 0.119831
J61107 0.065454
J61117 0.038518
J61127 0.087334
B0032 0.300000

The result of this experimental set up was a simple characterization of five of the Anderson RBS sequences in relation to other standard well-characterized RBS. The given relative strengths are displayed assuming B0034 as the unit.

We're also trying to look into mRNA folded shapes using mfold to see if there is a common pattern in the Anderson RBS shapes. This might be usable in predicting RBS strength for the other untested Anderson RBS sequences. Unfortunately, this seems not to be the case, as all the RBS sequences have very different mfold shapes.


Source data

Source code and data used for characterization:

Data: Experiment1_26_7_10.m

Matlab calculation code: IGEM_TUDelft_2010_Rbs_calc.m

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