Comparison of DNA extraction methods from archived samples for "long-read" sequencing techniques

Student thesis: Master's Thesis


The University of Faroe Islands and collaborating organizations set an aim to collect the information about genomes sequencing of animals inhabiting the territory of islands as well the costal water around the islands. For this study is necessary to collect the information about the genomes of vast kinds of species. To realize these purposes sequencing techniques with high – throughput and appropriate price are required.
Next generation sequencing technologies become more and more popular in scientific researches with their application to high – throughput analysis of biological systems. Isolation of pure, intact, double stranded, concentrated and not contaminated genomic DNA is prerequisite for successful and reliable large scale genetic analysis. In the present study, numbers of different DNA extraction techniques were examined to estimate their effectiveness for intact DNA extraction from preserved tissues of different fish species and whale. Four commercial DNA extraction kits as well in house developed methods were applied. The quality and quantity of the differentially extracted DNA was subsequently assessed by spectrometric measurements and gel electrophoresis. Extraction results revealed significant differences in quantity and quality of obtained DNA. Among applied techniques only heat induced methods showed its potential to extract DNA in demand quality from diverse samples. DNA obtained with other DNA extraction approaches had signs of degradation or was highly contaminated with proteins or chemical agents and did not meet requirements of set standards for the next generation sequencing. Furthermore, it was experimentally determined, that treated otoliths (which could be a source for historical DNA) are not appropriate as a reliable source of the DNA due to their acellular structure.
Date of Award2017
Original languageEnglish
Awarding Institution
  • Bremerhaven University of Applied Sciences
SupervisorSvein-Ole Mikalsen (Supervisor) & Stefan J. Wittke (Supervisor)

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