AbstractsChemistry

Synthesis of DNA-base selective labels for efficient labeling of DNA for TEM Imaging

by Rakesh Kumar




Institution: The University of Montana
Department: Chemistry
Degree: MA
Year: 2014
Keywords: Electrom Microscopy; TEM; Sequencing; Labelling; Platinum; DNA; Osmium; Cluster
Record ID: 2050412
Full text PDF: http://etd.lib.umt.edu/theses/available/etd-12032013-221432/


Abstract

The goal of this project was to explore the possibility of high throughput DNA sequencing using high-resolution electron microscopy. This approach was recently made feasible by development of a DNA threading technique that allows the imaging of straight, single stranded DNA; and by the development of atom-by-atom analysis with Transmission Electron Microscopy (TEM). In an attempt to better understand the types of interactions that can occur between water-soluble versions of complex and DNA we have synthesized the series of single metal and tetrametallic compounds. In order to demonstrate proof of concept for the technique of visualizing metal complex bound on DNA with TEM we studied the reaction of the complexes [Pt(en)Cl(NH2R)]+NO3- (en = ethylene diamine, Pt, Z = 78) with dGMP. We report here the successful visualization of the Pt atoms in the complex [Pt(en)Cl(NH2R)]+NO3- (R= C8H9NO2 (benzo[d][1,3]dioxol-5ylmethanamine, also known as piperonylamine) bound to DNA by TEM as well as kinetic study of a series of related complexes reacting with dGMP. The synthesis and dGMP binding of a Pt-triosmium conjugate is also reported.