Genomics Research Centre, Centre for Genomics and Personalised Health, Queensland University of Technology, Kelvin Grove, QLD, Australia.
Cancer Molecular Pathology of School of Medicine and Dentistry, Menzies Health Institute Queensland, Griffith University, Gold Coast, QLD, Australia.
Methods Mol Biol. 2022;2534:149-159. doi: 10.1007/978-1-0716-2505-7_11.
Single nucleotide polymorphisms (SNPs) can have a variety of implications for the progression and development of papillary thyroid carcinomas (PTCs). Identification of SNPs, either as germline variants or mutations occurring in tumor tissue, can thus have useful implications for patient management. There are many potential methods that can be used to identify a specific SNP or other genetic variant, and among these is high-resolution melting (HRM). HRM can be used to detect the presence of a genetic variant in a single sealed tube, involving undertaking a polymerase chain reaction (PCR) in the presence of a saturating intercalating dye. Once PCR is complete, the amplicons produced can be melted through incremental raising of the temperature and the genotype of individual samples determined by changes in the change in fluorescence as the fluorescent dye is released by the melting DNA. In this chapter, we detail a method for the genotyping of DNA samples using HRM.
单核苷酸多态性 (SNPs) 可能对甲状腺乳头状癌 (PTC) 的发展和进展有多种影响。因此，鉴定 SNPs（无论是种系变异还是肿瘤组织中发生的突变）对患者管理都有重要意义。有许多潜在的方法可用于鉴定特定的 SNP 或其他遗传变异，其中包括高分辨率熔解 (HRM)。HRM 可用于在单个密封管中检测遗传变异的存在，涉及在饱和嵌入染料存在下进行聚合酶链反应 (PCR)。一旦 PCR 完成，可以通过逐步升高温度来使扩增子解链，并通过荧光染料从解链 DNA 中释放时荧光的变化来确定单个样本的基因型。在本章中，我们详细介绍了使用 HRM 对 DNA 样本进行基因分型的方法。
