Difference between revisions of "Part:BBa K566004:Experience"
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===Experience=== | ===Experience=== | ||
| − | Testing and characterization for composite K566004 and K566006 in E. | + | Testing and characterization for composite [https://parts.igem.org/wiki/index.php?title=Part:BBa_K566004 K566004] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K566004 K566006] in ''E. coli'' JT2 strain under different light intensities and irradiation times of 30 minutes at 532 nm. |
Experimental | Experimental | ||
| − | G (+) = E.coli JT2 | + | G (+) = E.coli JT2 carrying pJT122, pPLPCB(S), K566004 (CFP), K566006 (Green light genetic inversor) |
| − | G (-) = E.coli JT2 | + | G (-) = E.coli JT2 carrying pJT122, K566004(CFP), K566006 (Green light genetic inversor) |
A light irradiation experiment was carried out in JT2 cultures under two controlled environments, one exposed to light and the other in complete darkness. The samples G (+) and G (-) were incubated overnight at 37°C and then 200uL of the samples were inoculated by triplicate for each intensity. | A light irradiation experiment was carried out in JT2 cultures under two controlled environments, one exposed to light and the other in complete darkness. The samples G (+) and G (-) were incubated overnight at 37°C and then 200uL of the samples were inoculated by triplicate for each intensity. | ||
| − | Both samples were measured in a Fluorometer BioTek Synergy, using special filters for CFP detection and measurement (Exitation filter 420nm ±50 and Emsion filter 460 ±40) before being irradiated to observe an initial fluorescence. After been introduced to the Light Machine, where the exposure time started with 30 minutes of Far-red light(710 nm) to reset the photoreceptors that could be switched during the night at the end of this period the fluorescence was measured. Then began to expose the samples to green light (532 nm) for 2 hours and every 30 minutes a portion of the sample is taken to measure the fluorescence | + | Both samples were measured in a Fluorometer BioTek Synergy, using special filters for CFP detection and measurement (Exitation filter 420nm ±50 and Emsion filter 460 ±40) before being irradiated to observe an initial fluorescence. After been introduced to the Light Machine, where the exposure time started with 30 minutes of Far-red light(710 nm) to reset the photoreceptors that could be switched during the night at the end of this period the fluorescence was measured. Then began to expose the samples to green light (532 nm) for 2 hours and every 30 minutes a portion of the sample is taken to measure the fluorescence gain or lost. |
[[Image:CFPgraficauno.png|thumb|center|500px|]] | [[Image:CFPgraficauno.png|thumb|center|500px|]] | ||
Revision as of 04:11, 29 September 2011
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Experience
Testing and characterization for composite K566004 and K566006 in E. coli JT2 strain under different light intensities and irradiation times of 30 minutes at 532 nm.
Experimental
G (+) = E.coli JT2 carrying pJT122, pPLPCB(S), K566004 (CFP), K566006 (Green light genetic inversor)
G (-) = E.coli JT2 carrying pJT122, K566004(CFP), K566006 (Green light genetic inversor)
A light irradiation experiment was carried out in JT2 cultures under two controlled environments, one exposed to light and the other in complete darkness. The samples G (+) and G (-) were incubated overnight at 37°C and then 200uL of the samples were inoculated by triplicate for each intensity.
Both samples were measured in a Fluorometer BioTek Synergy, using special filters for CFP detection and measurement (Exitation filter 420nm ±50 and Emsion filter 460 ±40) before being irradiated to observe an initial fluorescence. After been introduced to the Light Machine, where the exposure time started with 30 minutes of Far-red light(710 nm) to reset the photoreceptors that could be switched during the night at the end of this period the fluorescence was measured. Then began to expose the samples to green light (532 nm) for 2 hours and every 30 minutes a portion of the sample is taken to measure the fluorescence gain or lost.
Results
At the end of two hours the following data was obtained:
Plate Control (-)
- Always in the dark
User Reviews
UNIQ27ad58eef68389ac-partinfo-00000000-QINU UNIQ27ad58eef68389ac-partinfo-00000001-QINU