Yellow crucian carp high-density SNP (single nucleotide polymorphism) marker screening method and application
A screening method and high-density technology, applied in the field of molecular biology, can solve the problems of low purity of target DNA and complicated sequencing procedures, and achieve the effects of reducing library construction costs, consistent amplification efficiency, and simplifying complex genomes
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Embodiment 1
[0030] The high-density SNP marker screening method of yellow crucian carp comprises the following steps:
[0031] Genomic DNA extraction: Cut the crucian carp muscle or fin ray tissue sample into pieces and place in extraction solution I: 70ml of Tris-HCl and disodium hydrogen phosphate with a concentration of 120mM, 0.3g of fatty alcohol ether ammonium sulfate, and 30ml of chlorine with a concentration of 120mM Sodium chloride solution; add 0.5 μg plasmid DNA and 0.1 μg chitin, freeze and thaw 2-3 times, the freezing temperature is -166°C, the melting temperature is 50°C, centrifuge, collect the supernatant and add extract II: 10ml Tris-HCl with a concentration of 100mM and disodium hydrogen phosphate and 30ml of cationic polyacrylamide with a concentration of 0.5ppm were mixed upside down, centrifuged to produce a white precipitate, and the precipitate was washed to obtain a DNA sample for use; circular DNA and chitin The special existence, on the one hand, can promote the ...
Embodiment 2
[0038] The high-density SNP marker screening method of yellow crucian carp comprises the following steps:
[0039] Genomic DNA extraction: Mince the crucian carp muscle or fin ray tissue sample and place in extraction solution I: 70ml of Tris-HCl and disodium hydrogen phosphate with a concentration of 150mM, 0.5g of fatty alcohol ether ammonium sulfate, and 30ml of chlorine with a concentration of 130mM Sodium chloride solution; add 0.5 μg plasmid DNA and 0.1 μg chitin, freeze and thaw 2-3 times, the freezing temperature is -166°C, the melting temperature is 50°C, centrifuge, collect the supernatant and add extract II: 10ml Tris-HCl with a concentration of 100mM and disodium hydrogen phosphate and 30ml of cationic polyacrylamide with a concentration of 1.6ppm were mixed by inversion, centrifuged to produce a white precipitate, and the precipitate was washed to obtain a DNA sample for use;
[0040] RAD library construction and high-throughput sequencing: the extracted genomic D...
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