In English

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

Alexander Radenkovic
Göteborg : Chalmers tekniska högskola, 2017. 61 s.
[Examensarbete på avancerad nivå]

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.

Nyckelord: plasmids, deposinator, molecular combing, Cas9, S1 nuclease

Publikationen registrerades 2017-05-02. Den ändrades senast 2017-08-02

CPL ID: 249050

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