I. Pre-Phase One Experiments
Establish control experiments
§ -Constitutive GFP
§ o Switch H GFP
II. Pre-Phase One Goals
§ The goal of pre-phase 1 is to establish a working positive control and negative control in lab.
III. Phase One Experiments
§ Clone composite circuit in e.coli
§ Test efficacy of the circuit in vivo and determine threshold limits of both arsenic and arsenite
§ Perform mini prep and quantify
IV. Phase One Goals
§ The goals of this phase is to grow a large amount of the composite circuit and develop an understanding of whether or not the circuit is biologically effective. Another primary goal is to develop an understanding of how the circuit works in its entirety (upper threshold/lower threshold)
V. Phase Two Experiments
§ Test the efficacy of the composite circuit instead of crude lysate
§ Test purified plasmid with Promega S30 kit
VI. Phase Two Goals
§ The goal of this phase is to take the circuit out of the cell and test it in situ utilizing both a bacterial expression kit (Promega S30) and an attempt at functionalizing the circuit utilizing a formulated crude lysate for increased application space.
VII. Phase Three Experiments
§ Attempt at micro scaling reaction
§ Porting reactions to paper based system
§ Attempt at lyophilizing the circuit on paper both with Promega S30 kit and in the crude lysate form (if the previous phase demonstrated reasonable evidence for progressing on that front)
§ Phase Three Goals
§ The goal of this phase is to successfully micro scale the reaction and port it to a lateral flow paper based system. If successful in this phase, we will have developed a biological means to detect presence of arsenic and arsenite utilizing a lateral flow paper system that is both cost effective and easy to implement.
