Evaluation of novel cofactors for Methyltransferases labeling

The human genome is a complete set of genetic instructions that defines who we are at a molecular level. Comprising over 3 billion DNA base pairs, it encodes the information necessary for building and maintaining the human body. Comprehending the human genome is essential for advancing in fields such as genetics, medicine, and biotechnology, as it is pivotal to identifying genetic disorders, understanding disease mechanisms, and developing targeted treatments. In fact, over the past 47 years, since the early attempts in DNA sequencing, we have already witnessed multiple technological revolutions and growth in scale from a few kilobases to the first human genome, and now to millions of humans and a myriad of other genomes. It's inside this topic that we can collocate a novel technique that utilizes DNA methyltransferases to label specific sequences of DNA. In this project we tried to expand and investigate the current knowledge on cofactor and enzyme compatibility, used to achieve this Sequence-specific enzymatic DNA labeling. Chapter 1 discusses the literary background of DNA, microbiome and various methods to DNA sequencing. It begins with the definition of DNA and a brief explanation of its structure, moves on to the explanation of the microbiome and the various techniques developed over the years to study it, and concludes with an introduction to the purpose of this project. Chapter 2 is to introduce and explain the materials and methods used to execute our experiments. Starting from the first preliminary studies, to then conclude of the last steps of the analysis. Chapter 3 resumes all the experimental work done during this thesis project. At the end of this chapter new possible future developments for this technique are described, based on the findings achieved during this project. Supplementary material, including failed experiments and washing steps, is placed at the end of this thesis through the appendix referred to in Chapter 3.