Objective DNA analysis will become an important clinical tool in identification the composition of microbial species living in the gut. The most dominant technique at the time is based on PCR amplification of the 16s rDNA gene and subsequent sequencing of the variable region in the gene. This method however is limited in the amount of detail it provides and is notoriously susceptible to primer bias. Furthermore, due to the requirement of a PCR machine and sequencer the technique is too expensive and time consuming for a clinical setting. Over the past few years nanopores have been heralded as a cheap alternative for biomolecular analysis. Specifically solid-state nanopores, which consist of a small pore in a solid membrane, have the potential for cost-effective mass-production in cleanroom facilities. Furthermore, since these pores can detect double stranded DNA they can easily be used for analysis of DNA maps.In this proposal I suggest to use DNA mapping with solid state nanopores. The basic idea is to sequence-specifically label DNA molecules using DNA Methyltransferases and synthetic analogues of the natural cofactor S-adenosyl-methionine. The pattern of labels attached to the DNA is unique for the underlying sequence.In my Ph.D. I used this method to transfer fluorescent dyes to the DNA and subsequently extracted the DNA map using super resolution fluorescence microscopy. In this case, the attached label will induce a current-drop when the DNA molecule translocates through the channel. The extracted current-trace will then be converted to a DNA map in basepairs and can be used to match to a real sequence. This proposed method can be used to identify genomic elements and eventually be used to recognize species, for a fraction of the cost of 16s rDNA sequencing. Fields of science natural sciencesbiological sciencesgeneticsDNAnatural sciencesphysical sciencesopticsmicroscopynatural sciencesbiological sciencesmicrobiology Programme(s) H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions Main Programme H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility Topic(s) MSCA-IF-2016 - Individual Fellowships Call for proposal H2020-MSCA-IF-2016 See other projects for this call Funding Scheme MSCA-IF-EF-ST - Standard EF Coordinator ECOLE POLYTECHNIQUE FEDERALE DE LAUSANNE Net EU contribution € 175 419,60 Address Batiment ce 3316 station 1 1015 Lausanne Switzerland See on map Region Schweiz/Suisse/Svizzera Région lémanique Vaud Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Other funding € 0,00
