Fluorescently-labeled insertion/deletion (InDel) genetic polymorphism locus composite amplification system and application thereof
A composite amplification system and a technology of the composite amplification system are applied in the field of biological detection to achieve the effects of high polymorphism, rapid amplification technology and accurate typing results
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[0051] Example 1 Application of InDel Site Fluorescence Multiplex Amplification System for Paternity Identification and Individual Identification
[0052] Steps:
[0053] 1. DNA extraction
[0054] Genomic DNA extraction by Chelex-100 method: add 1mL double-distilled water to a 1.5mL centrifuge tube, then add 3μL whole blood or 3×3mm blood spot, mix carefully; incubate at room temperature for 30min, shake intermittently; centrifuge at 14,000 r / min for 5min; Carefully remove the supernatant. Add 200 μL of 10% Chelex 100 (100mesh) to the pellet; incubate at 56°C for 30 min; shake at high speed for 5-10 sec; bath in boiling water for 8 min; centrifuge at 14,000 r / min for 5 min, and take the supernatant for PCR amplification.
[0055] 2. PCR amplification
[0056] The total reaction volume was 15.0 μL, including 7.5 μL of PCR reaction buffer (Buffer), 3.0 μL of primer mix (Primer pair Mix), 0.5 μL of DNA polymerase, 3.0 μL of template DNA, and 1 μL of ultrapure water.
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