Team:Bielefeld-Germany/Protocols/Analytics
From 2012.igem.org
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The [http://www.sigmaaldrich.com/catalog/product/sigma/53739?lang=de®ion=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. | The [http://www.sigmaaldrich.com/catalog/product/sigma/53739?lang=de®ion=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. | ||
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. | 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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{{Team:Bielefeld/Sponsoren}} | {{Team:Bielefeld/Sponsoren}} |
Revision as of 09:16, 13 September 2012
Activity measurements
For the measurements [http://www.sigmaaldrich.com/catalog/product/sigma/m0312?lang=de®ion=DE 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 |
[http://www.sigmaaldrich.com/catalog/product/sigma/a1888?lang=de®ion=DE ABTS] | 0,1 mM |
H2O | ad 200 µL |
The [http://www.sigmaaldrich.com/catalog/product/sigma/53739?lang=de®ion=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 10-5 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.
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