Abstract
The article presents the results of a comparison of isolation methods during SNP genotyping of cattle. Successful genotyping depends on obtaining DNA from samples in sufficient quantity and quality to carry out molecular reactions. Three methods of nucleic acid extraction were used in the work. The method of isolation using a commercial PureLink Genomic DNA Mini Kit (Thermo Fisher Scientific, USA) showed the highest degree of yield and purity of nucleic acids. When DNA was isolated from the hair follicles of cattle with the PureLink Genomic DNA Mini Kit, the average DNA concentration in the samples was 151.78 ng/ml, DNA extraction with the DNA-sorb-B kit was 21.51 ng/ml and the phenol-chloroform method corresponded to 175.08 ng/ml. The extracted animal DNA was later used for genotyping by the SNP method. Most of the evaluated extraction methods allowed amplification of a significant amount of DNA. The results of genotyping in samples isolated using PureLink Genomic DNA Mini Kit compared to other samples showed 100% amplification of products. Of the methods tested in this study, extraction with this set was the most effective in terms of further DNA detection.
01 Introduction
The full text of the article is available for download in PDF format on the right panel.
02 References
- 1. Столповский Ю.А. Популяционно-генетические основы сохранения генофондов доместицированных видов животных. Вавиловский журнал генетики и селекции, 2013, т.17, № 4/2, С.901-213.
- 2. Манцевич Е.А., Епишко О.А., Генотипирование крупного рогатого скота по гену LEP. УО «Гродненский государственный аграрный университет», г. Гродно, Республика Беларусь, 2019 г.,С. 146-150.
- 3. Ганджа А.И., Курак О.П., Леткевич Л.Л., Симоненко В.П., Кириллова И.В., Журина Н.В., Ковальчук М.А. // Генотипирование эмбрионов крупного рогатого скота на основе ДНК-анализа. РУП «Научно-практический центр Национальной академии наук Беларуси по животноводству», г. Жодино, Минская обл., Республика Беларусь, 2014.
- 4. Янич Ф., Влияние полисахаридов на продуктивные качества новотельных коров голштино-фризской породы в условиях тоо «БЕК+». Izdenister Natigeler, (2 (94), С.12–20. 2022 https://doi.org/10.37884/2-2022/02.
- 5. Эрнст Л.К., Дмитриев Н.Г., Паронян И.А., Истомин А.А. Генетические ресурсы сельскохозяйственных животных в России и сопредельных государствах. СПб., 1994, С.473.
- 6. Моисеева И.Г., Уханов С.В., Столповский Ю.А. и др. Генофонды сельскохозяйственных животных: генетические ресурсы животноводства России/Под ред. И.А. Захарова. М.:Наука, 2006. С. 466.
- 7. Эрнст Л.К., Зиновьева Н.А. Биологические проблемы животноводства в ХХ1 веке. М.,2008. С.505.
- 8. Fernández, María E. Comparison of the effectiveness of microsatellites and SNP panels for genetic identification, traceability and assessment of parentage in an inbred Angus herd // Genetics and Molecular Biology 36, 2, P. 185 – 191 2013.
- 9. Barker K. Phenol-Chloroform Isoamyl Alcohol (PCI) DNA Extraction / Barker K // At the bench: a laboratory navigator. 1st edn. (Cold Spring Harbor Laboratory Press, New York, 1998) P. 284–289.
- 10. Deniskova, T.E., Sermyagin, A.A., Bagirov, V.A. et al. Comparative analysis of the effectiveness of STR and SNP markers for intraspecific and interspecific differentiation of the genus Ovis . Russ J Genet 52, P. 79–84 (2016).
- 11. Morin, P.A., Luikart, G., and Wayne, R.K., The SNP workshop group: SNPs in ecology, evolution and con servation, Trends Ecol. Evol., 2004, vol. 19, P. 208– 216. 7.
- 12.Vignal, A., Milan, D., Sancristobal, M., and Eggen, A., A review on SNP and other types of molecular markers and their use in animal genetics, Genet. Sel. Evol., 2002, vol. 34, P. 275–305.