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

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dc.contributor.authorRadenkovic, Alexander
dc.contributor.departmentChalmers tekniska högskola / Institutionen för biologi och biotekniksv
dc.contributor.departmentChalmers University of Technology / Department of Biology and Biological Engineeringen
dc.date.accessioned2019-07-03T14:27:33Z
dc.date.available2019-07-03T14:27:33Z
dc.date.issued2017
dc.description.abstractAntibiotic 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.
dc.identifier.urihttps://hdl.handle.net/20.500.12380/249050
dc.language.isoeng
dc.setspec.uppsokLifeEarthScience
dc.subjectBiologiska vetenskaper
dc.subjectBioinformatik och systembiologi
dc.subjectLivsvetenskaper
dc.subjectBiological Sciences
dc.subjectBioinformatics and Systems Biology
dc.subjectLife Science
dc.titleEstimation of Plasmid Lengths on ZEONEX®-Coated Cover Slips Using a Deposinator for Stretching of Single DNA Molecules
dc.type.degreeExamensarbete för masterexamensv
dc.type.degreeMaster Thesisen
dc.type.uppsokH
local.programmeBiotechnology (MPBIO), MSc
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