Difference between revisions of "Part:BBa K574009:Experience"
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'''Methods:''' we ligated E0840 (''<partinfo>E0840</partinfo>'') downstream of K574009, and the positive colony was cultured at 37°C, 220 rpm for 12 hours, as well as the DH5alpha. Then we diluted 1:25 the former in 12 falcon tubes (4 ml cultures), each 3 tubes as a group. The later was also diluted in 3 tubes as the negative control. After Cultured for another 2 hours, the groups were induced with 3OC12HSL 0, 10^-6M, 10^-5M and 10^-4M respectively. We took samples after 1 hours, 4 hours, 8 hours and 24 hours, and then the samples were measured through the flow cytometer. | '''Methods:''' we ligated E0840 (''<partinfo>E0840</partinfo>'') downstream of K574009, and the positive colony was cultured at 37°C, 220 rpm for 12 hours, as well as the DH5alpha. Then we diluted 1:25 the former in 12 falcon tubes (4 ml cultures), each 3 tubes as a group. The later was also diluted in 3 tubes as the negative control. After Cultured for another 2 hours, the groups were induced with 3OC12HSL 0, 10^-6M, 10^-5M and 10^-4M respectively. We took samples after 1 hours, 4 hours, 8 hours and 24 hours, and then the samples were measured through the flow cytometer. | ||
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'''Results:''' From the chart, we can see that cells were hardly induced in the control group, and with the concentration of inducer growing, the intensity of GFP increased by groups. The most efficient concentration of inducer was around 10^-6M, and to most groups, the intensity of GFP reached its maxium after 4 hours. | '''Results:''' From the chart, we can see that cells were hardly induced in the control group, and with the concentration of inducer growing, the intensity of GFP increased by groups. The most efficient concentration of inducer was around 10^-6M, and to most groups, the intensity of GFP reached its maxium after 4 hours. | ||
Revision as of 16:52, 4 October 2011
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Applications of BBa_K574009
Description: we assembled E0840 (BBa_E0840) under the pLas promoter (BBa_R0079) that was contained into K574009. We kept pSB1A2 as the scaffold vector.
Methods: we ligated E0840 (BBa_E0840) downstream of K574009, and the positive colony was cultured at 37°C, 220 rpm for 12 hours, as well as the DH5alpha. Then we diluted 1:25 the former in 12 falcon tubes (4 ml cultures), each 3 tubes as a group. The later was also diluted in 3 tubes as the negative control. After Cultured for another 2 hours, the groups were induced with 3OC12HSL 0, 10^-6M, 10^-5M and 10^-4M respectively. We took samples after 1 hours, 4 hours, 8 hours and 24 hours, and then the samples were measured through the flow cytometer.
Results: From the chart, we can see that cells were hardly induced in the control group, and with the concentration of inducer growing, the intensity of GFP increased by groups. The most efficient concentration of inducer was around 10^-6M, and to most groups, the intensity of GFP reached its maxium after 4 hours.
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