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Kit used for detecting body size traits of chicken and molecular breeding method of chicken
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A molecular breeding and kit technology, applied in the field of molecular genetics, can solve the problems of long time, many influencing factors, and complicated SSCP operation, and achieve the effect of increasing the width of the pelvis and the length of the body oblique.
Inactive Publication Date: 2015-10-07
NANYANG NORMAL UNIV
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However, SSCP is cumbersome, time-consuming, has many influencing factors, and there are false negative problems in the experimental process, so it is not an ideal SNP detection method; the cost of direct sequencing technology is relatively high
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Embodiment 1
[0035] In the present embodiment, the kit for detecting chicken body size traits includes:
[0036] 10pmol / μl upstream primer F PRimer: 5'-ACGTTGGATGTTCTGCCATACCAGTGTGTG-3',
[0037] 10pmol / μl downstream primer R PRimer: 5'-ACGTTGGATGACTCCTCCAGCTGCTGCTTC-3';
[0040]In this embodiment, the kit used to detect the body size traits of chickens, in addition to the following primers, also includes: 10 × buffer, 25mM Mg 2+ , 25mM dNTP, 5U / μl Hotstar, the reaction concentration is 1×iPLEX single-base elongase, SAP buffer, 1.7U / μl SAP enzyme, ddd H 2 O and DNA Marker;
[0041] 10pmol / μl upstream primer F PRimer: 5'-ACGTTGGATGTTCTGCCATACCAGTGTGTG-3',
[0042] 10pmol / μl downstream primer R PRimer: 5'-ACGTTGGATGACTCCTCCAGCTGCTGCTTC-3';
[0131] The molecular breeding method based on chicken miRNA-1658 gene SNP in the present embodiment may further comprise the steps:
[0132] 1) Extraction of total DNA and PCR amplification
[0133] The peripheral blood DNA of 4-week-old chickens was extracted by the phenol-chloroform-isoamyl alcohol method, dissolved in an appropriate amount of TE (pH 8.0), stored at -20°C for later use, and PCR-amplified with primers F Primer / R PRimer to obtain the amplified product;
[0134] Upstream primer F PRimer: 5'-ACGTTGGATGTTCTGCCATACCAGTGTGTG-3',
[0135] Downstream primer R PRimer: 5'-ACGTTGGATGACTCCTCCAGCTGCTGCTTC-3';
[0136] The reaction system of PCR amplification is: total 5.0μl; 10×buffer 0.5μl, 25mM Mg 2+0.4μl, 25mM dNTP 0.1μl, 5U / μl Hotstar 0.2μl, 10pmol / μl F Primer / R Primer 0.5μl each, 100ng / μl total DNA 1.0μl, ddd H 2 O 1.8 μl;
[0137] The reaction program of PCR amplification is: pre-denaturation at 95°C for 2min; 95°C for 30s; 56°C for 30s; 72°C for 60s, 45 cycles;...
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Abstract
The invention discloses a kit used for detecting body size traits of chicken and a molecular breeding method of the chicken and belongs to the field of molecular genetics. MassARRAY system matrix-assisted laserdesorption / ionization time-of-flight mass spectrometry and a single-primer extension reaction are combined, miRNA (micro ribonucleic acid)-1658 (rs16681031 C / G) single-SNP (single nucleotide polymorphism) site genotyping detection on F2 generation resource population samples of Gushi chicken and Anak chicken is performed, and correlation analysis with the size traits of F2 generation resource populations is performed. Results show that the chicken miRNA-1658 gene SNP (single nucleotide polymorphism) is obviously correlated with the body size traits of the chicken; a GG gene facilitates obvious increasing of chest breadth, crista breadth and body slanting length of a four-week-year-old chick; chicken flocks with excellent genetic resources can be quickly built by taking the GG gene as assistant selection of the body size trait of the four-week-year-old chick and as a molecular genetic breeding marker of the four-week-year-old chick.
Description
technical field [0001] The invention relates to a kit for detecting chicken body size traits, and also relates to a molecular breeding method based on chicken miRNA-1658 gene single nucleotide polymorphism, which belongs to the field of molecular genetics. Background technique [0002] With the rapid development of social economy and the improvement of people's living standards, the market's demand for meat quality has generally shifted from the previous quantity requirement to the pursuit of quality and flavor. Growth traits, body size traits, meat quality traits and carcass traits are the most economical traits in the poultry industry today. Most economic traits are quantitative traits, which are mainly determined by a large number of micro-effect genes. These genes have additive effects, dominant effects and epistasis effects at the same time. The genetic basis of complex traits is the genetic factors that affect the phenotype. It is jointly determined by environmental f...
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