Team:Imperial College/3HB assay

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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.
<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 19:20, 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. <p>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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