Researcher(s)
- Thea Tantral-Johnson, Pre-Veterinary Medicine and Animal Biosciences, University of Delaware
Faculty Mentor(s)
- Dr. Behnam Abasht, Animal and Food Sciences, University of Delaware
Abstract
In the US, poultry is an $81.7 billion industry and is a major protein source for millions of people. Increases in meat production have come alongside increases in growth-related myopathies. Specifically, Wooden Breast Disease (WBD) is a condition affecting the pectoralis (p.) major muscle of chickens, causing myodegeneration and fibrosis, which lowers meat quality. While the disease causes significant economic losses for producers, the exact causes and mechanisms of the myopathy are still unknown. This project aims to investigate the relationship between genetic polymorphisms and histopathologic tissue features for the purpose of expanding knowledge on the genetic basis of WB. Primers were designed using NCBI Primer-BLAST then tested for long range amplification of 4-5kb regions spanning the promoter and extending into the gene body of selected candidate genes from the Gallus gallus broiler chicken reference genome (GRCg7b; RefSeq Accession: GCF_016699485.2). A subset of 50 primer pairs were selected based on product strength on gel electrophoresis from 212 primers tested. These selected primers were grouped into 5 pools based on thermodynamic compatibility and minimization of cross-primer dimers using ThermoFisher’s Multiple Primer Analyzer. Ultimately, once primer pooling for multiplex PCR has been optimized, products will go through DNA library preparation using Oxford Nanopore’s Rapid Barcoding Kit 96 v14, then sequencing using a MinIONMk1D sequencer. This will generate amplicon reads that can be analyzed for polymorphisms as well as how these polymorphisms relate to physical changes in muscle tissue. Overall, the goal of this project is to identify genes strongly associated with histological WBD characteristics while also informing primer design and pooling practices for future work.