Assessing genetic potential of lambs involves taking blood from jugular vein from animals, performing PCR and sheep genotyping, amplifying, electrophorizing, visualizing and calculating frequency of occurrence of alleles and genotypes
	KARPOVA E D; SUROV A I
	2023-08-06
专利权人	NORTH CAUCASIAN FEDERAL SCI AGRARIAN CEN (NCAU-Non-standard)
申请日期	2023-08-06
专利号	RU2023120552-A; RU2835927-C2
成果简介	NOVELTY - Method for assessing genetic potential of lambs based on gene analysis involves taking blood from jugular vein from animals as biomaterial, performing PCR and sheep genotyping with respect to the GH, CAST and GDF9 gene complex, amplifying DNA fragment on a programmable four-channel thermal cycler in a volume of 20 μ l comprising 17 μ l reaction mixture and 3 μ l DNA, 15 μ l reaction mixture and 5 μ l DNA using the primers of SEQ ID NOs: 1 and 2, 3 and 4, and 5 and 6, after which a restriction analysis of the obtained amplifications is performed using restriction endonucleases HaeIII, MspI, BstHH1, performing electrophoresis in 2-4% agarose gel stained with ethidium bromide, placing on a transilluminator filter emitting light in the UV range, visualizing number and lengths of restriction fragments, establishing presence and frequency of alleles and genotypes, determining calculation of the frequency of occurrence of genotypes and calculating frequency of occurrence of alleles. USE - The method is useful for assessing genetic potential of lambs based on gene analysis. DETAILED DESCRIPTION - Method for assessing genetic potential of lambs based on gene analysis by amplifying a gene fragment using primers, preferably growth hormone (GH) gene having 277 base pair (bp) using forward primer comprising a base pair sequence of SEQ ID NO: 1 and reverse primer comprising a base pair sequence of SEQ ID NO: 2, calpastatin (CAST) having 622 bp using forward primer comprising a base pair sequence of SEQ ID NO: 3 and reverse primer comprising a base pair sequence of SEQ ID NO: 4 and growth differentiation factor 9 (GDF9) having 462 bp using forward primer comprising a base pair sequence of SEQ ID NO: 5 and reverse primer comprising a base pair sequence of SEQ ID NO: 6, involves taking blood from jugular vein from animals as biomaterial, performing PCR and sheep genotyping with respect to the GH, CAST and GDF9 gene complex, amplifying DNA fragment on a programmable four-channel thermal cycler in a volume of 20 μ l comprising 17 μ l reaction mixture and 3 μ l DNA, 15 μ l reaction mixture and 5 μ l DNA using the above-mentioned primers, after which a restriction analysis of the obtained amplifications is performed using restriction endonucleases HaeIII, MspI, BstHH1, performing electrophoresis in 2-4% agarose gel stained with ethidium bromide, placing on a transilluminator filter emitting light in the UV range, visualizing number and lengths of restriction fragments, using standard M 50 'GenePakDNAMarkers' set as a molecular mass marker, establishing presence and frequency of alleles and genotypes, determining calculation of the frequency of occurrence of genotypes using specific equation involving predetermined parameters such as p, n and N, where p is frequency of a certain genotype, n is number of animals with a certain genotype and N is total number of animals, and calculating frequency of occurrence of alleles using specific equation involving predetermined parameters such as p, n and N, where P is frequency of occurrence of the allele, A is allele, N1 is number of homozygotes for the allele under study, N2 is the number of heterozygotes and n is sample size of animals.
IPC 分类号	A01K-067/02 ; C12Q-001/68
国家	俄罗斯
专业领域	生物科学
语种	英语
成果类型	专利
文献类型	科技成果
条目标识符	http://119.78.100.226:8889/handle/3KE4DYBR/20956
专题	中国科学院新疆生态与地理研究所
作者单位	NORTH CAUCASIAN FEDERAL SCI AGRARIAN CEN (NCAU-Non-standard)
推荐引用方式 GB/T 7714	KARPOVA E D,SUROV A I. Assessing genetic potential of lambs involves taking blood from jugular vein from animals, performing PCR and sheep genotyping, amplifying, electrophorizing, visualizing and calculating frequency of occurrence of alleles and genotypes. RU2023120552-A; RU2835927-C2[P]. 2023.

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