Achievements

Policies and Practices:

• Thoroughly researched our target markets and differing legislations that could affect our system
• Connected with many different experts in the fields of infectious diseases, anti-malarial medications and public health to truly understand the needs and feasibility of the project
• Connected with the foremost authority in low cost diagnostics, Foundation for Innovative New Diagnostics (FIND), to discuss the viability of our design and concept
• Connected with a small village clinic in rural Uganda, which gave an opportunity to see the facility as well as discuss the topic with local physicians
• Featured iGEM and our project on a global stage at the United Nations, in Geneva, and created discussion regarding the advancement of synthetic biology

Modelling:

• Designed and manufactured a prototype of informed design based on our biological system
• Cost analysis of our prototype and system to ensure that this project is feasible in developing nations
• Quantified the visible threshold of detection for several reporters using time-lapsed cell count and absorbance readings, as well as a programmed colour sensor
• Programmed an Arduino UNO microcontroller, colour sensor, and LCD screen to detect colour change among detection chambers, allowing for quantification of the colour
• Applied circuitry of a heating pad/temperature considerations for ideal isothermal PCR conditions
• Used Autodesk Maya Software to visually model and represent our system
• Used Solid-works Software to model fluid flow analysis for different prototype designs

Transformation:

• Transformed B. subtilis with a primer product containing 125 bp of homologous sequence flanking our gene of interest
• Modified and optimized general transformation: characterized effect of changing xylose concentrations, temperature, and shaking
• Optimization of transformation in small, liquid volume
• Sporulated B. subtilis
• Activated B. subtilis spores back to vegetative state
• Isolated the master regular for competency, comK from the genome of B. subtilis using PCR and inserted it into a Biobrick vector for future application as an iGEM standard part.

Sample Preparation:

• Demonstrated efficacy of isothermal PCR to amplify DNA sequences without the need of a thermocycler, using freeze-dried components
• Demonstrated ability to amplify target DNA sequence from whole E. coli cells in a blood mixture
• Obtained specificity in target sequence primers - specific to numerous strains of N. meningititis but not other pathogens including a commensal strain of N. meningititis

Detection/Genetic circuit

• Designed composite regulatory promoter sequences - used to allow repression of a downstream reporter with a specific repressor. Includes 5 variants of repressible promoters in either single or triplicate repeats for variation in strength of repression
• Demonstrated interaction (i.e. repression) between the reporter gene and repressor gene in E. coli
• Demonstrated the efficacy of homologous recombination in B. subtilis and showed the minimum length of flanking sequences
• Biobricked and submitted the full-length lacZ gene, previously non-existent in the latest distribution plates
• Constructed a reporter operon consisting of a constitutive promoter (Pveg), ribosome binding site, operator (C1434) and reporter (LacZ/RFP).
• Constructed a complementary repressor operon consisting of a constitutive promoter (Pveg), ribosome binding site, and repressor pertaining to the operator. Repressor was also flanked with regions homologous to certain sequences within our pathogens of interest.