Direct TaqMan-PCR based technology for non-invasive genotyping of lipid metabolism associated SNPs
10.16352/j.issn.1001-6325.2024.06.0858
- VernacularTitle:基于直接TaqMan-PCR的无创检测脂质代谢相关SNP基因分型技术
- Author:
Yuanhong SUN
1
;
Lyu XIE
;
Lihua FAN
;
Zhi ZHENG
Author Information
1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 生物化学与分子生物学系,北京 100005
- Keywords:
direct PCR;
nucleic acids extraction;
SNP genotyping;
high-throughput
- From:
Basic & Clinical Medicine
2024;44(6):858-865
- CountryChina
- Language:Chinese
-
Abstract:
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 stor-age 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 homo-zygous wild type,homozygous mutant type and heterozygote at rs688 and rs964184 loci.Conclusions A fast,con-venient,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.
