Team:Bielefeld-Germany/Protocols/Analytics

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=List of all other protocols we use in our project=
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==Activity measurements ==
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For the measurements [http://www.sigmaaldrich.com/catalog/product/sigma/m0312?lang=de&region=DE 96-well flat bottom microplates] were used. Each well contained a total sample volume of 200 µL respectively.
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The sample setup was pipetted as follows:
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{| class="wikitable"
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|-
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! component !! concentration
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|-
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| buffer: || 100 mM
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|-
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| Laccase || 0,1 U
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|-
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| [http://www.sigmaaldrich.com/catalog/product/sigma/a1888?lang=de&region=DE ABTS] || 0,1 mM
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|-
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| H<sub>2</sub>O || ad 200 µL
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|}
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The [http://www.sigmaaldrich.com/catalog/product/sigma/53739?lang=de&region=DE Laccase of ''T. versicolor''] we bought for standardization was diluted in water so that 140 µL would contain 0,1 U meaning 72 x 10<sup>-5</sup> g of the enzyme. Please check the [https://2012.igem.org/Team:Bielefeld-Germany/Labjournal labjournal] if Sodium-Acetate or [https://2012.igem.org/Team:Bielefeld-Germany/Protocols#Briton_Robinson_Buffer Briton Robinson Buffer] was used, respectively.
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The ABTS and laccase concentration optimum for standardization (so that the reaction was traceable nicely) was determinated during several experiments. Check the [https://2012.igem.org/Team:Bielefeld-Germany/Labjournal labjournal] for further information.
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Revision as of 09:06, 13 September 2012

List of all other protocols we use in our project

Activity measurements

For the measurements 96-well flat bottom microplates were used. Each well contained a total sample volume of 200 µL respectively. The sample setup was pipetted as follows:

component concentration
buffer: 100 mM
Laccase 0,1 U
ABTS 0,1 mM
H2O ad 200 µL

The Laccase of T. versicolor we bought for standardization was diluted in water so that 140 µL would contain 0,1 U meaning 72 x 10-5 g of the enzyme. Please check the labjournal if Sodium-Acetate or Briton Robinson Buffer was used, respectively. The ABTS and laccase concentration optimum for standardization (so that the reaction was traceable nicely) was determinated during several experiments. Check the labjournal for further information.

55px Logo merck.jpg BioCircle.JPG Bielefeld2012 Evonik.jpg Bielefeld2012 Baxter.png Logo knauer.jpg Logo iit.jpg Bielefeld2012 BIEKUBA.jpg Logo biometra.jpg Logo bio-nrw.png Bielefeld2012 Logo ERASynbio.jpg



55px Logo merck.jpg BioCircle.JPG Bielefeld2012 Evonik.jpg Bielefeld2012 Baxter.png Logo knauer.jpg Logo iit.jpg Bielefeld2012 BIEKUBA.jpg Logo biometra.jpg Logo bio-nrw.png Bielefeld2012 Logo ERASynbio.jpg