Difference between revisions of "Part:BBa K902002:Experience"

 
 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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how you used this part and how it worked out.
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===Applications of BBa_K902002===
 
===Applications of BBa_K902002===
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===User Reviews===
 
===User Reviews===
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<partinfo>BBa_K902002 AddReview 5</partinfo>
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<I>Calgary 2012</I>
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This biobrick was used as a novel electrochemical reporter by the Calgary 2012 iGEM team. It has been shown to cleave the chemical para-nitrophenol-&beta;-D-glucuronide (PNPG) into glucuronic acid and para-nitrophenol (PNP). The resultant phenol, PNP, oxidizes at a voltage of 1.6V vs the Reduction of Hydrogen Electrode (RHE), allowing for electrochemical detection of the level of the chemical present in the solution.
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Detection of the product of the cleavage of PNPG into PNP was carried out at a carbon working electrode in a solution of 25mL 0.1M pH7 PBS. This was done with or without the addition of 100&micro;M IPTG to induce the <i>lacI</i> promoter. While detection is observed in the uninduced sample due to leaky expression, the levels of induced expression is much higher almost immediately after induction.
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[[Image:Calgary2012 ECHEM PNP.png‎|thumb|600px|center|Figure 1: Potentiostatic detection of PNP at 1.6V vs RHE by either an IPTG induced <i>uidA</i> gene or leaky expression of an uninduced <i>uidA</i> gene.]]
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Latest revision as of 19:39, 3 October 2012


Applications of BBa_K902002

User Reviews

UNIQ5d40020a6c14f825-partinfo-00000000-QINU UNIQ5d40020a6c14f825-partinfo-00000001-QINU

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Calgary 2012

This biobrick was used as a novel electrochemical reporter by the Calgary 2012 iGEM team. It has been shown to cleave the chemical para-nitrophenol-β-D-glucuronide (PNPG) into glucuronic acid and para-nitrophenol (PNP). The resultant phenol, PNP, oxidizes at a voltage of 1.6V vs the Reduction of Hydrogen Electrode (RHE), allowing for electrochemical detection of the level of the chemical present in the solution.

Detection of the product of the cleavage of PNPG into PNP was carried out at a carbon working electrode in a solution of 25mL 0.1M pH7 PBS. This was done with or without the addition of 100µM IPTG to induce the lacI promoter. While detection is observed in the uninduced sample due to leaky expression, the levels of induced expression is much higher almost immediately after induction.

Figure 1: Potentiostatic detection of PNP at 1.6V vs RHE by either an IPTG induced uidA gene or leaky expression of an uninduced uidA gene.