Jichu yixue yu linchuang (Jun 2024)

Direct TaqMan-PCR based technology for non-invasive genotyping of lipid metabolism associated SNPs

  • SUN Yuanhong, XIE Lyu, FAN Lihua, ZHENG Zhi

DOI
https://doi.org/10.16352/j.issn.1001-6325.2024.06.0858
Journal volume & issue
Vol. 44, no. 6
pp. 858 – 865

Abstract

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Objective To establish a non-invasive single nucleotide polymorphism genotyping technology based on direct TaqMan-PCR for high-throughput genotyping of site rs688 and rs964184 associated with lipid metabolism to meet the need for early screening of at-risk populations. Methods Extracted DNA was used to optimize the PCR annealing extension temperature and amplification procedure for the designed TaqMan probe; the oral swabs were placed into the sample treatment solution and then briefly centrifuged, and a small amount of the supernatant was taken for the direct qPCR amplification analysis; the freeze-thawing stability of probe and the effects of sample storage time on genotyping results were explored. Results The technology requires only simple treatment of oral swab for PCR analysis and can be stored at room temperature for 4 d. The optimized method was able to distinguish homozygous wild type, homozygous mutant type and heterozygote at rs688 and rs964184 loci. Conclusions A fast, convenient, high-throughput, and low-cost SNP genotyping method has been established, providing an efficient and accurate new approach for large-scale screening of high-risk populations carrying susceptibility genes.

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