Method for simultaneously detecting 32 single nucleotide polymorphism (SNP) locus genotypes
A single nucleotide polymorphism, gene technology, applied in the field of medical molecular biology, can solve the problems of poor sensitivity, easy missed diagnosis, misdiagnosis, difficult to popularize and so on
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[0069] Example 1 The genotypes of 32 single nucleotide polymorphism sites in the Chinese Han population were simultaneously detected by the method of the present invention.
[0070] (1) Designing 32 pairs of primers and 32 extended primers for sequencing and hybridization for amplifying fragments containing the target single nucleotide polymorphism site, see sequence 1 to sequence 96 for details. .
[0071] (2) Dilute and mix the PCR primers so that the final concentration is 2.5 μM, construct the PCR primer pool and amplify the target fragment. The PCR reaction system for every 96 people is:
[0072]
[0073] Reagent Components Volume (μL)
[0074]
[0075] Primer Pool (2.5μM) 11.2
[0076] Deoxyribonucleotides (2.5mM) 20
[0077] 10x PCR Buffer II 56.2
[0078] Magnesium chloride (25mM) 112.6
[0079] Gold Amplification Enzyme (5U / μL) 11.2
[0080] Double distilled water 126
[0081] ...
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