A method for identification of different migratory populations of Suifenhe three blockfish
A group and migration technology, applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve problems such as germplasm mixing and morphological discrimination errors, and achieve simple steps, strong specificity, and identification The results are accurate
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specific Embodiment approach 1
[0023] Specific embodiment 1: This embodiment is used to identify the microsatellite primer pair of Sankuai fish, wherein the black beach head effective detection marker microsatellite primer pair is SSR-1, and the nucleotide sequence of the SSR-1 forward identification primer is 5'- CTTGTGGTGCTGGTTCTTCA-3'; the nucleotide sequence of the SSR-1 reverse identification primer is 5'-GATGTGGCAGACCCTGACTT-3'; the golden beach head effective detection marker microsatellite primer pair is SSR-2, and the SSR-2 forward identification primer nucleotide The sequence is 5'-TGGTTGGACAGCAAACAAAG-3'; the nucleotide sequence of the SSR-2 reverse identification primer is 5'-TGAAGGTCTGGCTGATGATG-3'.
specific Embodiment approach 2
[0024] Specific implementation mode two: the method for identification of different migratory groups of Suifenhe three-piece fish in this embodiment is carried out according to the following steps:
[0025] 1. Extraction of sample DNA;
[0026] 2. PCR detection:
[0027] The PCR reaction system is 15 μl, consisting of 10.8 μl self-made mixed buffer, 0.5 μl forward detection primer, 0.5 μl reverse detection primer, 2 μl sample DNA, 0.2 μl Taq DNA polymerase with an enzyme activity of 1U and 1 μl deionized sterile water ; Among them, 0.5 μl of forward identification primer contains 10 mmol / L of forward identification primer, 0.5 μl of reverse identification primer contains 10 mmol / L of reverse identification primer, 2 μl of sample DNA contains 50 ng of sample DNA obtained in step 1, and 10.8 μl of homemade The mixed buffer contains 50mmol / L of KCl, 10mmol / L of Tris-HCl, 0.1% volume of TritonX-100, 1.5mmol / LM of MgCl 2 , 0.1% by volume of NP-40, 0.01% by volume of gelatin and 2...
specific Embodiment approach 3
[0036]Specific embodiment 3: The difference between this embodiment and specific embodiment 2 is: Step 1: Take the sample fish fin and place it in a 1.5ml centrifuge tube, then add 600 μl of lysate and 10 μl of proteinase K in sequence, mix well and place at 55°C Digest overnight in a water bath; after digestion, add 10 μl RNase and mix well, then place in a water bath at 37°C for 1 hour; then add 600 μl of a mixture of phenol and chloroform, mix well, and centrifuge at 12,000 r / min for 10 minutes at room temperature; take the centrifuged supernatant Place in another empty centrifuge tube, add 1ml of absolute alcohol and mix well, centrifuge at 12000r / min for 10min at room temperature; after centrifugation, spin to pour out the alcohol, then add 500μl of 70% ethanol, and centrifuge at 12000r / min for 5min at room temperature; Then use a pipette gun to remove the ethanol, and then centrifuge the residue at 12000r / min for 1min. After the centrifugation, pour out the residual alcoh...
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