From 2014.igem.org
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| - | <i>Escherichia coli</i> is the model organism in which a system is implemented that is able to trigger a reaction in and of this organism to the presence of compounds leeching from landmines. <i>Escherichia coli</i> carries promoters of genes hypothesized to respond to compounds containing nitrogen. DNT, TNT and DBP contain this element and leech from landmines. By coupling of fluorescent molecule mKATE2 to (improved versions of) these promoters, qualification of the response and hence of the presence of the compound is possible. This module thus encompasses the development of a biosensor.
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| - | Modeled are the responses of <i>Escherichia coli</i> to compounds DNT, TNT and DNB in a straightforward fashion. Input are compounds, generated is the response of the promoter; datapoints resulting from imaging mKATE2 fluorescence and linking fluorescence to compounds added at defined points in time after induction with relevant compounds. Black box in this Module are the regulatory enzymes, possibly nitrogen-linked, and genes encoding these elements.
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| - | Experimental work centers on responsive promoters, compounds, and, in addition, response depending on several enzymes functional in nitrogen metabolism in order to shine light on the black box regarding the reaction and actual molecular basis of reaction to compounds leeching from landmines.
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| - | Implementation of a system of microfluidics within this Module is based on the aspiration to be able to perform in-field measurements of compounds leeching from landmines. The necessity of having a certain type of device points towards the use of microfluidics. Aspects central in development of this type of device are, amongst others, the speed of reactions, the possibility of in vivo measurements at all points in time and options for quantification.
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| - | Aim, construction and functionality of what is hereafter referred to as the mother machine are discussed under [.]. This type of microfluidic system is intended for single-cell experiments, utilizing nanoscale channels coupled to a central channel for flow of media. Team iGEM 2014 TU Delft has made use of the standard model developed at Massachusetts Institute of Technology.
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| - | Summarizing, the device is constructed via positive silicon waver coupled to a plastic mold. The team has constructed its negative with PDMS, generating channels. Size of channels and size of cells should be taken in consideration, the latter partly depending on growth conditions. Holes and tubes coupled to these holes, coupled to the central channel deal with flushing with medium, with respect to the Module Landmine Detection carrying compounds DNT, TNT or DNB. Before application, the PDMS sample is connected to a glass slide by oxygenplasma and prepared for action by consecutive flushings with, amongst others, BSA. [imagen] More information regarding the design and construction of the mother machine can be found in the section [.].
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Revision as of 13:21, 16 October 2014
Integration of Departments
click to return to the Module Landmine Detection
Described are the general why and how of the practices Modeling as well as Microfluidics with respect to experimental work performed in the module Landmine Detection.
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