Part:BBa_K2722003:Experience
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how you used this part and how it worked out.
Applications of BBa_K2722003
We used this part to create a composite part, which expresses this mTQ protein. This fluorescent protein was used to quality control the expression efficiency of our self-made cell extract in comparison to purchased cell extracts. Cell extract is a composition of all E.Coli plasma proteins, that has all the required machinery for transcription and translation processes outside of a living cell. This can be used for rapid characterisation of biorbicks, genetic circuits, expression of toxic proteins and a lot more.
We have measured mTQ fluorescence at different concentrations. Giving a linear concentration to fluorescence blot, which might be used to calculate concentrations from fluorescence.
The extinction coefficient ε is calculated with following formula: ε=A/(c ∙l)
Whereby A is the Absorption, c is the concentration in Mol and l is the pathlength of the probe. We measured the Absorption at 434 nm wavelength of six samples. Three having a concentration of 22 µM and another three having a concentration of 11 µM mTQ protein on a Nanophotometer (l = 1 mm). And calculated the extinction coefficient.
Absorption Extinction coefficient [1/Mcm]
Sample1: 22 µM;Absorption:0,068; Extinction coefficient: 30,909 1/Mcm
Sample2: 22 µM Absorption:0,076 Extinction coefficient:34,545 1/Mcm
Sample3: 22 µM Absorption:0,075 Extinction coefficient:34,091 1/Mcm
Sample4: 11 µM Absorption:0,031 Extinction coefficient:28,182 1/Mcm
Sample5: 11 µM Absorption:0,030 Extinction coefficient:27,273 1/Mcm
Sample6: 11 µM Absorption:0,030 Extinction coefficient:27,273 1/Mcm
Mean extinction coefficient value: 30,379 1/Mcm
We generated a mean value from our six samples of 30,379 1/Mcm. Which is consistent with the literature value.
User Reviews
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