A new approach for DNA sequencing by End-Labelled Free-Solution Electrophoresis (ELFSE) is proposed. The focus of the work is directed toward the design of novel end-label molecules that can be uniquely attached to DNA strands in order to separate DNA in the absence of a polymer matrix. The successful implementation of ELFSE as a practical and rapid approach for DNA separation will supplant current gel-based sequencing methods by decreasing run-time and increasing data output. The postdoctoral fellow will explore the use of biopolymeric materials of linear and branched architectures as basis for end-labels. In vitro and in vivo approaches will be used to generate linear polypeptides. Branched end-labels will be produced by grafting linear polypeptides with synthetic polypeptoids, more specifically N-substituted glycines. The postdoctoral fellow will use liquid chromatography, NMR spectroscopy. mass spectrometry, and circular dichroism to determine the identity, level of monodispersity, and structural conformation of the biomolecules. The chemical composition, length and structure of the end-labels will be optimized for efficiently separating DNA fragments in free solution. For achieving highly efficient separation of long DNA sequencing fragments (>600 bases) by capillary electrophoresis, the postdoctoral fellow will optimize capillary length and coating, electric field intensity, buffer conditions, and injection method..
