Difference between revisions of "Part:BBa K784000:Experience"

(→‎Experience of BBa_K784000)
(→‎Experience of BBa_K784000)
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We have used the part inside a BL21 strain of ''E.coli'' which has an endogenouse PET system. <br>
 
We have used the part inside a BL21 strain of ''E.coli'' which has an endogenouse PET system. <br>
 
In the presence of IPTG inducer, T7 RNAP is expressed and activates the part promoter which expresses the Alkaline Phosphatase gene (phoA).<br>
 
In the presence of IPTG inducer, T7 RNAP is expressed and activates the part promoter which expresses the Alkaline Phosphatase gene (phoA).<br>
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We checked the absorbance resulted from the [https://parts.igem.org/Arkin_JCA_PNPAssay  PNP (para-nitrophenol phosphate) hydrolysis assay] in raising concetrations of the inducer (IPTG).The error bars represent the processing of the data collected. The line at the bottom of the graph represents the basal level according to the control result- no IPTG induction.The results are before you in the graph: <br>
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The graph below presents the results of the experiment described. The absorbance at 420nm for ‎each of the different concentrations of the inducer in a range of 0.5 μΜ to 1000 μΜ on a ‎logarithmic scale. The absorbance was calculated via different dilutions of the samples: dilutions ‎by 0.5, by 0.25, and by 0.1. The error bars represent the processing of the data collected. The line at ‎the bottom of the graph represents the basal level according to the control result- no IPTG ‎induction.‎ <br>
  
 
[[Image:technionphoagraph1.jpg|300X300px|frame|center]]
 
[[Image:technionphoagraph1.jpg|300X300px|frame|center]]
  
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As you can see in the graph, raising concentrations of the inducer results in raising absorbance, which means raising concentration of the Alkaline Phosphatase enzyme.
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As can be seen, the graph shows a clear positive tendency- the higher the concentration the higher ‎the absorption, as expected. Starting from a concentration of 40 μM and above, there are only ‎small deviations from the absorption value of  1.2, probably due to the fact that saturation has ‎been achieved. ‎
  
 
===User Reviews===
 
===User Reviews===

Revision as of 22:16, 24 September 2012

Experience of BBa_K784000

We have used the part inside a BL21 strain of E.coli which has an endogenouse PET system.
In the presence of IPTG inducer, T7 RNAP is expressed and activates the part promoter which expresses the Alkaline Phosphatase gene (phoA).
The graph below presents the results of the experiment described. The absorbance at 420nm for ‎each of the different concentrations of the inducer in a range of 0.5 μΜ to 1000 μΜ on a ‎logarithmic scale. The absorbance was calculated via different dilutions of the samples: dilutions ‎by 0.5, by 0.25, and by 0.1. The error bars represent the processing of the data collected. The line at ‎the bottom of the graph represents the basal level according to the control result- no IPTG ‎induction.‎

300X300px

As can be seen, the graph shows a clear positive tendency- the higher the concentration the higher ‎the absorption, as expected. Starting from a concentration of 40 μM and above, there are only ‎small deviations from the absorption value of 1.2, probably due to the fact that saturation has ‎been achieved. ‎

User Reviews

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