Part:BBa_K1157006:Experience

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Applications of BBa_K1157006

We used flow cytometry to test the individual fluorescence expression of our biobrick from 0 to 4 hours and in different concentration(10^-3, 10^-5,10^-7).

User Reviews

UNIQ9384a15d532ed1bc-partinfo-00000000-QINU

• iGEM_Stockholm_2015

Method: Promoter, RBS, RhlI, and double terminator BioBricks were transformed in TOP10 E. coli using the transformation protocol and were supposed to be assembled using the 3A assembly protocol. Results: During the initial lab weeks there were some trouble with transformation, later addressed to not using KCM buffer. The BBa_K082035 was ordered from IDT to speed up the process after repeated failed transformations and transformation troubleshooting were done in parallel. Conclusion: BBa_K082035 was ordered instead of cloned. Please, visit our [http://2015.igem.org/Team:Stockholm iGEM Stockholm 2015 webpage] for more details or contact us.

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•• iGEM_Stockholm_2015

Method: E. coli transformed with BBa_K1157006 were grown together with bacteria transformed with BBa_K082035 on normal agar plates without antibiotics. Results: The plates showed a slight red color change showed in Figure below. Figure: Plate showing E. coli with BBa_K082035 spread in horizontal streaks E. coli with BBa_1157006 spread as an M on top. A slight red color change is seen, as expected. Conclusion: Even though the two plates showed the correct color change, the results were however not reproducible at a later stage. Please, visit our [http://2015.igem.org/Team:Stockholm iGEM Stockholm 2015 webpage] for more details or contact us.

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••• iGEM_Stockholm_2015

Method: For each experiment fresh solutions in was prepared in ddH2O in the following concentrations: 100 mM; 10 mM; 0.1 mM; 0.01 mM. We tested these different concentrations in 3 repetitions first in agar plates with the bacteria with BBa_K1157006. Then we tested these different concentrations in the liquid LB cultures with 3 repetitions for each sample. Both plates and liquid cultures were incubated in 37°C for 24h. Results: No color change was observed for any of the experiments. It seems that our bacterial strain did not respond to any concentrations of BHL in agar or liquid cultures. Conclusion: Based on our results, this part of our experiment did not work as intended. Please, visit our [http://2015.igem.org/Team:Stockholm iGEM Stockholm 2015 webpage] for more details or contact us.

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