期刊文献+

焦磷酸测序分析CTR1基因多态性方法的建立

Development of pyrosequencing method for detection of CTR1 polymorphism
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摘要 目的:建立CTR1 rs10981694单核苷酸多态性位点的焦磷酸测序方法。方法:全血g DNA为模板,生物素标记引物,对扩增多态位点目的片段,制备生物素标记单链模板,与测序引物退火结合后行焦磷酸测序。分析结果经毛细管电泳测序验证,并进行重复性检验。结果:本文建立了针对CTR1 rs10981694多态性位点的焦磷酸测序方法,经毛细管电泳测序验证和重复性验证,结果准确可靠。CTR1 rs10981694 C和A等位基因频率分别为21%和79%均符合Hardy-Weinberg平衡。结论:本文建立的焦磷酸测序方法可准确、高通量、快速检测CTR1 rs10981694单核苷酸多态性,并且特别适宜大样本量的临床及科研批量检测需要。 AIM: To establish a pyrosequencing based method for detection of CTR1 rs10981694 polymorphisms and of the frequency of these polymorphisms in healthy Chinese. METHODS: The target fragments were amplified by PCR from 193 healthy subjects'gDNA samples, polymorphisms were detected on PyroMark ID by pyrosequencing technology. The reliability of pyrosequencing methods were validated by repeat tests and Sanger sequencing. RESULTS:A new pyrosequencing method was established to detect the CTR1 rs10981694 polymorphisms in healthy Chinese. The detection rate and repetition rate were both 100%. The allele frequencies of CTR1 rs10981694 C and A were 21% and 79% respectively. Genotype frequencies match the Hardy-Weinberg equilibrium. CONCLUSION: The pyrosequencing assay for detecting CTR1 polymorphisms is proved to be a rapid, accurate and high- throughput methods, and it can be a preferred alternative to conventional methods in research and clinical application.
出处 《中国临床药理学与治疗学》 CAS CSCD 2016年第6期673-677,共5页 Chinese Journal of Clinical Pharmacology and Therapeutics
基金 国家自然科学基金项目(81402968) 中央高校基本科研业务费(2012QNZT085 2012QNZT133)
关键词 药理学 焦磷酸测序 CTR1 单核苷酸多态性 pharmacology pyrosquencing CTR1 SNP
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