Estimation of Plasmid Lengths on ZEONEX®-Coated Cover Slips Using a Deposinator for Stretching of Single DNA Molecules

Examensarbete för masterexamen

Please use this identifier to cite or link to this item: https://hdl.handle.net/20.500.12380/249050
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Type: Examensarbete för masterexamen
Master Thesis
Title: Estimation of Plasmid Lengths on ZEONEX®-Coated Cover Slips Using a Deposinator for Stretching of Single DNA Molecules
Authors: Radenkovic, Alexander
Abstract: Antibiotic resistance is one of the largest threats to global health, as it counters the way we treat bacterial infections. Antibiotic resistance genes can spread be-tween bacteria via plasmids. Simple and efficient methods for studying plasmids are thus important in the global fight against antibiotic resistance. This project optimizes and evaluates a novel molecular combing method for length estimation of plasmids – dragging a droplet of DNA solution across a ZEONEX®-coated cover slip followed by fluorescence microscopy. The method was optimized for pH, deposition speed and concentration using -DNA. A plasmid containing the resistance gene blaCTX−M−15 was linearized using either Cas9 or S1 nuclease, and the efficiency of both endonucleases was assessed. The plasmid was accurately estimated to 150 kbp by using -DNA as reference. The method shows promise, but further optimization is required to reduce noise that arise from broken DNA fragments.
Keywords: Biologiska vetenskaper;Bioinformatik och systembiologi;Livsvetenskaper;Biological Sciences;Bioinformatics and Systems Biology;Life Science
Issue Date: 2017
Publisher: Chalmers tekniska högskola / Institutionen för biologi och bioteknik
Chalmers University of Technology / Department of Biology and Biological Engineering
URI: https://hdl.handle.net/20.500.12380/249050
Collection:Examensarbeten för masterexamen // Master Theses



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