Difference between revisions of "Part:BBa K1157006:Experience"

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<partinfo>BBa_K1157006 AddReview 1</partinfo>
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<I>iGEM_Stockholm_2015</I>
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'''Method:''' Promoter, RBS, RhlI, and double terminator BioBricks were transformed in TOP10 ''E. coli'' using the [https://static.igem.org/mediawiki/2015/c/cc/STHLM_Transformation_protocol.pdf transformation protocol] and were supposed to be assembled using the [https://static.igem.org/mediawiki/2015/f/f6/STHLM_3A_assembly_protocol.pdf 3A assembly protocol].
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'''Results:''' During the initial lab weeks there were some trouble with transformation, later addressed to not using KCM buffer. The [https://parts.igem.org/Part:BBa_K082035 BBa_K082035] was ordered from IDT to speed up the process after repeated failed transformations and transformation troubleshooting were done in parallel.
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'''Conclusion:''' [https://parts.igem.org/Part:BBa_K082035 BBa_K082035] was ordered instead of cloned.
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Please, visit our [http://2015.igem.org/Team:Stockholm iGEM Stockholm 2015 webpage] for more details or contact us.
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<I>iGEM_Stockholm_2015</I>
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'''Method:''' ''E. coli'' transformed with BBa_K1157006 were grown together with bacteria transformed with [https://parts.igem.org/Part:BBa_K082035 BBa_K082035] on normal agar plates without antibiotics.
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'''Results:''' The plates showed a slight red color change showed in Figure below.
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[[File:STHLM_readout_oneplate.png]]
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''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.''
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'''Conclusion:''' Even though the two plates showed the correct color change, the results were however not reproducible at a later stage.
 +
 
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Please, visit our [http://2015.igem.org/Team:Stockholm iGEM Stockholm 2015 webpage] for more details or contact us.
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|};
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<partinfo>BBa_K1157006 AddReview 3</partinfo>
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<I>iGEM_Stockholm_2015</I>
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'''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.
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'''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.
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'''Conclusion:''' Based on our results, this part of our experiment did not work as intended.
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Please, visit our [http://2015.igem.org/Team:Stockholm iGEM Stockholm 2015 webpage] for more details or contact us.
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|};

Latest revision as of 15:36, 18 September 2015

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.

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). Flow mCherry.jpg

User Reviews

UNIQdb7b1e16616fb2ed-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.

;
••

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.

STHLM readout oneplate.png 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.

;
•••

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.

;