Difference between revisions of "Part:BBa K875001:Experience"
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| + | To test this promoter we performed two different assays. | ||
| + | First of all we cloned this promoter in pSB1C3 and then we inserted downstream the GFP BBa_I13504. In the same plasmid we cloned J23100-CymR-B0015 in order to repress the promoter. To test this system we used different concentrations of cumate which binds CymR preventing its repression, thus allowing the GFP expression. | ||
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | ||
how you used this part and how it worked out. | how you used this part and how it worked out. | ||
Revision as of 13:42, 23 September 2012
To test this promoter we performed two different assays.
First of all we cloned this promoter in pSB1C3 and then we inserted downstream the GFP BBa_I13504. In the same plasmid we cloned J23100-CymR-B0015 in order to repress the promoter. To test this system we used different concentrations of cumate which binds CymR preventing its repression, thus allowing the GFP expression.
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Trieste Team 2012
In liquid assay:
We inoculated a 20ml culture. After overnight growth, we diluted the culture to OD600 = 0.2. Then we aliquoted 200 μl in 8 replicates in a microtiter plate at different concentrations of cumate. The reading was performed in a monochromator at 485-510nm.
In the plate assay:
We streaked the culture on LB Agar plates containing different p-cumate concentrations.
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