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Team:BCCS-Bristol/Wetlab/BetaGalactosidaseAssays
From 2010.igem.org
Contents |
β-Galactosidase Assays
Motivation
In order to properly map the PyeaR part in the range of soil nitrate concentrations that agriColi would be exposed to, we needed to get some data on the sensitivity of the component within that small range of concentrations. To that end, we decided that a β-galactosidase assay would be the most appropriate method of obtaining that data, especially given that Edinburgh were kind enough to provide us with the necessary biobrick to run the tests.
Unfortunately it turned out that the [http://partsregistry.org/Part:BBa_K216009 BBa_K216009] part was not entirely appropriate for the assay - the data could be limited by the cellular expression of the LacZΩ portion of β-galactosidase which is under the regulation of a separate promoter on the chromosomal DNA. So we decided to build our own biobrick to enable us to run the tests (see the page on design of the PyeaR+LacZ construct for more details). However this wasn't a success and so we returned to [http://partsregistry.org/Part:BBa_K216009 BBa_K216009] in order to get data on the PyeaR part.
The assay
The general procedure for the assay, whether using the [http://partsregistry.org/Part:BBa_K216009 BBa_K216009] part or our own biobrick (we made one attempt to use our own construct before realising that the ligation had failed).
