Evaluation of the effectiveness of DNA extraction methods from sheep ear tissue samples for whole-genome sequencing
DOI:
https://doi.org/10.51452/cajvs.2026.3(015).2248Keywords:
Whole genome sequencing, DNA extraction, sheep ear notches, STAB, PureLink Genomic DNA Kit, Kazakh fat-tailed sheep breedAbstract
Background and Aim. Whole-genome sequencing (WGS) is a powerful tool for studying the genetic structure of livestock populations. Effective implementation of WGS is largely determined by the quality of the source genomic DNA. Sheep ear notches obtained in the field are a convenient source of biomaterial; however, their high connective tissue density makes DNA extraction challenging. The aim of this study was to compare the STAB method and the commercial PureLink Genomic DNA Kit for DNA extraction from ear notches of Kazakh fat-tailed sheeps.
Materials and Methods. The study was conducted in two stages. STAB lysis buffer was used in the first stage, and the commercial PureLink Genomic DNA Kit (Thermo Fisher Scientific) was used in the second stage. Quality control was performed using a NanoDrop2000 spectrophotometer and gel electrophoresis.
Results. The average DNA concentration isolated by the CTAB method was 293 ng/μl, and the A260/280 ratio ranged from 1.70 to 1.86 (mean 1.78 ± 0.04), indicating heterogeneous purity. The PureLink kit demonstrated consistently high quality: concentration 350.2 ± 96.9 ng/μl, A260/280 1.86 ± 0.01, and A260/230 2.13 ± 0.08.
Conclusion. The commercial PureLink Genomic DNA Kit is recommended for DNA extraction from sheep ear notches, as it provides consistent quantitative and qualitative data that meet WGS requirements.
