Part:BBa_I746907:Experience

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User Reviews

TU_Munich Team 2011

We tested the part and it worked very well.

TU_Munich Team 2011 GFP Assay

I746907 (T7 promotor with GFP) in pSB1A2 in BL21DE3 cells

change of medium to M63 (LB shows slight fluorescence itself)

five concentrations of IPTG in the medium were tested (1.5, 1.0, 0.5, 0.25, 0.1 mM IPTG) growth of cells at 37 °C, shaking

excitation filter: 485 nm, emission filter: 520 nm

Problem: measurement of Absorbance only every 100 minutes led to spikes in the diagram (Fluorescence measured every 10 min.)

results

part works well

induction t(1/2) = ca. 200 min

no concentration dependency of GFP induction at 1.5 to 0.1 mM IPTG, as IPTG concentration was in saturation range

for this diagram:

• calculation: Fluorescence/Absorbance
• mean of triplicate measurement
• adjusted to 100 %
• corrected for outliers
• shortened to 700 min

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