Difference between revisions of "Part:BBa R0080:Experience"

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<I>Username</I>
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<I>Xinyuan Qiu NUDT_CHINA2014</I>
 
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This promoter is lack of AraC gene so that the promoter can only activate the down-stream expression with the existence of both AraC and Arabinose
  
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===Materials and Methods===
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We used the circuit of BBa_K1532008 to test the function of this part, the sequence and features of the part BBa_K1532008 is as follows:
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https://static.igem.org/mediawiki/2014/f/f9/NUDT_CHINA_2014_K1532008.png
  
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Fluorometric measurement were processed with Fluoroskan(R) Ascent FL from ThermoTM using the filters of 485nm and 538nm. The E.coli to be tested was cultured in liquid LB media containing 35μg/mL chloramphenicol to OD1.6, than split into 96 well plates (from ThermoTM) by 150μL/well. The plate was then put into the Fluoroskan(R) Ascent FL and cultured in 25℃ and background shake of 90rpms. The Fluorometric measurement was processed with the interval time of 10 minutes.
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The layout of the plate is as follows:
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https://static.igem.org/mediawiki/2014/5/50/NUDT_CHINA_R0080_CON1.jpg
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We draw the curve using the intensity figure of different amount of Arabinose subtracted by the figures of those without Arabinose.
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===Results===
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The result is as follows:
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https://static.igem.org/mediawiki/2014/8/89/NUDT_CHINA_2014_RCC80_CON2.jpg
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The results indicates that with the existence of Arabinose and AraC, the part BBa_R0080 can activate the down-stream expression observably.
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Latest revision as of 01:32, 19 October 2014

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_R0080

User Reviews

UNIQd5911b012e9132cb-partinfo-00000000-QINU

••

Xinyuan Qiu NUDT_CHINA2014

This promoter is lack of AraC gene so that the promoter can only activate the down-stream expression with the existence of both AraC and Arabinose

Materials and Methods

We used the circuit of BBa_K1532008 to test the function of this part, the sequence and features of the part BBa_K1532008 is as follows: NUDT_CHINA_2014_K1532008.png

Fluorometric measurement were processed with Fluoroskan(R) Ascent FL from ThermoTM using the filters of 485nm and 538nm. The E.coli to be tested was cultured in liquid LB media containing 35μg/mL chloramphenicol to OD1.6, than split into 96 well plates (from ThermoTM) by 150μL/well. The plate was then put into the Fluoroskan(R) Ascent FL and cultured in 25℃ and background shake of 90rpms. The Fluorometric measurement was processed with the interval time of 10 minutes.

The layout of the plate is as follows:

NUDT_CHINA_R0080_CON1.jpg


We draw the curve using the intensity figure of different amount of Arabinose subtracted by the figures of those without Arabinose.

Results

The result is as follows:

NUDT_CHINA_2014_RCC80_CON2.jpg

The results indicates that with the existence of Arabinose and AraC, the part BBa_R0080 can activate the down-stream expression observably.


;

Stephanie

The promoter is not inducible by arabinose as it does not contain the required araC gene.

Use I0500 instead which contains the araC gene and the Pbad arabinose controlled promoter.

;


iGEM TUDelft 2008, Ostassen

We've used this promoter in our project, and have also found it is always on, independent of arabinose. This result can be found [http://2008.igem.org/Team:TUDelft/Temperature_results#Setting_up_luciferase_measurements here] in figure 3, and has been confirmed on more occasions later.

;

UNIQd5911b012e9132cb-partinfo-00000004-QINU