Team:Imperial College/3HB assay

From 2013.igem.org

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<h1 class="clear">3HB assay</h1>
<h1 class="clear">3HB assay</h1>
<p><b>This experiment completes the cycle of PHB bioplastic recycling. </b></p>
<p><b>This experiment completes the cycle of PHB bioplastic recycling. </b></p>
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<p>Please see [https://2013.igem.org/Team:Imperial_College/Protocols#3HB_assay 3HB protocol] for sample preparation and the way we adapted the kit for our purposes.
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<p>Please see [https://2013.igem.org/Team:Imperial_College/Protocols#3HB_assay 3HB protocol] for sample preparation and the way we adapted the kit for our purposes.</p>
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<p>The well setup was as shown below:</p>
<p>The well setup was as shown below:</p>
https://static.igem.org/mediawiki/parts/4/40/96_well_plate_3HB.jpg
https://static.igem.org/mediawiki/parts/4/40/96_well_plate_3HB.jpg

Revision as of 19:22, 4 October 2013

3HB assay

This experiment completes the cycle of PHB bioplastic recycling.

Please see 3HB protocol for sample preparation and the way we adapted the kit for our purposes.

The well setup was as shown below:

96_well_plate_3HB.jpg

We prepared serveral dilutions of the samples and measured the absorbance at 445, 450 and 455.

Collected_data_for_3HB_assay.JPG

We corrected the absorbance values busubstracting the 0 standard.

Data_collection_3HB_assay2.JPG

We plotted the standard curve and used the below formula to calculate the 3HB concentrations in our samples.

Formula_for_calculation.JPG

MM_3HB_calculation.JPG

We have taken the dynamic range of the assay into account (according to protocol) and therefore excluded readings where the concentration of 3HB in the well was above 1 mM because these are not accurate.

3HB_from_PHB_from_waste.jpg

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