Hardware

Our dedicated effort revolves around expanding access to DNAzyme technology for detection, specifically by prioritizing the use of renewable materials and economic platforms. Through targeted educational initiatives and the integration of affordable technologies, we aim to make this innovative detection approach available in remote and economically challenged regions. The overarching objective is to democratize the knowledge and resources associated with DNAzyme applications, ensuring that individuals in financially constrained communities can effectively utilise renewable materials and cost-effective platforms for precise and affordable detection processes. This approach seeks to promote sustainability and affordability, fostering inclusivity in adopting DNAzyme technology across diverse socio-economic landscapes.

Test Strip

To achieve this goal, we decided not to use graphene oxide for the immobilisation of the DNAzyme, rather we used nitrocellulose, which is a cheaper material to produce. The design goals also kept modularity as a priority, when making the flow kits for DNAzyme detection. We decided to use a single strip divided into 3 segments by using graphite from pencils. This through experimentation was found to be an excellent substitution for other plastic/silicon-based channels. The design of the strip has been iteratively simplified to be as simple as possible and was finally reduced to just a paper with graphite channels.

PreRNA workflow

Aptamer generation from a random library takes around 15 SELEX cycle. The majority of the sequences do not bind very well to the target protein, due to non-complex folding. Most aptamers have multi-stem loop structures, which only appear after the 4th cycle of SELEX. To hasten this process we have developed the PreRNA workflow, which uses RNA library of a cell to generate aptamers. This is done, as most mRNAs have complex structures, which allow the to interact with several proteins. The methods are summarised as below:

We extracted the mRNA from HEK293 cell line, and started the SELEX cycle against e.coli DH5alpha. The results are sent for sequencing. We also have developed another workflow, to generate aptamers from gDNA, especially repeat regions. This mainly has the benefit of starting with the same stock of gDNA or Plasmid and doing an initial round of EP-PCR to generate variants of the PLasmids or gDNA. This is followed by blunt digestion and sonication to break apart into multiple short fragments. These are tagged with an unequal duplex, that have inbuilt priming sites that can then be used for aptamer generation, via asymmetric EP-PCR.