SNP locus related to growth characteristics of patinopecten yessoensis and detection and application thereof
A technology for growth traits and scallops, applied in the field of molecular breeding genetic marker screening, can solve the problems of unseen IGF system gene research
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Embodiment 1
[0022] The cDNA sequence cloning of the scallop IGFBP gene in the present invention comprises the following steps:
[0023] a) extraction of total RNA from adductor muscle of scallop scallop;
[0024] b) synthesis of cDNA first strand;
[0025] c) Obtaining the full-length cDNA sequence of the target gene;
[0026] d) Bioinformatics analysis of the target gene.
[0027] The specific operation is as follows:
[0028] a) Extraction of total RNA from Ezo scallop. Total RNA was extracted from adductor muscle of Ezo scallop according to the Trizol method.
[0029] b) Synthesis of cDNA first strand. Using 2μg total RNA as a template, add 1μl 20μM Oligo d(T) 18 , RNase&DNase-free H 2 O to make up to 13μl, mix and centrifuge. Incubate at 70°C for 10 min, and immediately place on ice to open the RNA secondary structure. Add in sequence: 5μl 5×M-MLV Buffer, 5μl dNTP (2.5mM), 1μl RNasin (40U / μl), 1μl M-MLV (200U / μl); mix well and centrifuge at 42°C for 90min. 94°C, 5min, to ina...
Embodiment 2
[0035] Example 2: Screening analysis of SNP sites related to growth traits in the IGFBP gene of Ezo scallop and its method for assisting high-yielding scallop breeding
[0036] The correlation analysis between the growth traits-related SNP site C.1054A>G and the growth traits in the IGFBP gene of the scallop scallop and the method for its application in the breeding of the high-yielding scallop include the following steps:
[0037] a) extraction of scallop genome DNA;
[0038] b) SNP site screening of the IGFBP gene of the scallop;
[0039] c) Design of primers and probes for locus C.1054A>G typing;
[0040] d) SNP C.1054A>G typing;
[0041] e) Correlation analysis between C.1054A>G genotype and growth traits;
[0042] f) The method of SNP C.1054A>G assisted breeding of high-yielding Chlamys farreri.
[0043] The specific operation is as follows:
[0044] a) Extraction of DNA from Ezo scallop. Take about 0.1 g of adductor muscle, add 500 μl of STE lysis buffer, cut it into...
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