Laura Plazer

06.12.2022

In situ mini DNA barcode sequencing and SNP typing in cheetahs (Acinonyx jubatus) using Oxford Nanopore Technologies (MinION)

MSc Student
Advisor: Elisabeth Haring

Natural History Museum &
Unit for Integrative Zoology, Department of Evolutionary Biology
University of Vienna

Abstract

Rapid advances in sequencing technologies provide us with a wide range of possibilities and the development of miniaturized laboratory equipment allows for genetic analyses in settings lacking laboratory infrastructure, funding or dealing with restrictions concerning transportation of samples.The Oxford Nanopore Technologies’ (ONT) MinION sequencer allows for rapid analyses in situ.Until recently the technology had some issues, such as a relatively high error rate compared to other high-throughput sequencing platforms, or the minimum read length of 200bp. However, in the last months ONT has made several improvements to their chemistry, flow cells and software tools. In this study we want to test if these improvements, coupled with ONT’s new Short Fragment Mode (SFM) allow us to carry out reliable SNP typing or DNA mini-barcode sequencing on the MinION. We do so using SNPs and mini-barcode amplicons developed for threatened cheetahs (Acinonyx jubatus). First results suggest that short sequences can be reliably called, opening new possibilities towards easy and affordable ways for evidence-based conservation methods as well as whole genome sequencing in situ.