Sekvensbaserad artbestämning av veterinärmedicinska prover

• Main Authors: Andersson, Louise, Solander, Klara, Brokmar, Linde, Bylander, Karl, Pölder, Magdalena, Henriksson, Nellie
• Format: Others
• Language: Swedish
• Published: Uppsala universitet, Institutionen för biologisk grundutbildning 2021
• Subjects: parasiter; primer; PCR; Natural Sciences; Naturvetenskap; Agricultural and Veterinary sciences; Lantbruksvetenskap och veterinärmedicin
• Online Access: http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-444228

In this project a PCR primer library was created to facilitate the amplification of DNA of parasites that are morphologically indistinguishable. The aim of this paper is to present a PCR primer library which is applicable to many of the species of interest, as well as creating a protocol for handling samples that contain more than one unknown species. The results presented in this rapport were achieved by means of literature studies as well as in silico primer design and validation. The literature studies focused mainly on finding pre-existing primers, whilst the phylogenetic studies retrieved information necessary for primer design. Each primer, whether achieved through literature or in silico assembly, was validated according to certain predefined criterion. The finalized PCR primer library contains 96 primers covering a total of 789 parasites. Beyond this, multiple primers that did not meet our criterion are presented. This due to promising results from the studies in which these were used. An ethical analysis of treatment against parasites was conducted. Finally, how to use the library is discussed containing relevant methods that could be helpful to further develop a detailed protocol. Methods such as Sanger sequencing, Next-generation sequencing and gel electrophoresis are in focus.