数字聚合酶链反应
核酸
小RNA
分子生物学
核糖核酸
计算生物学
化学
荧光
实时聚合酶链反应
聚合酶链反应
生物
生物化学
基因
物理
量子力学
作者
Jiayi Peng,Feng Li,Xiangdong Xu,Shen Hu
出处
期刊:Methods in molecular biology
日期:2023-01-01
卷期号:: 169-177
被引量:1
标识
DOI:10.1007/978-1-0716-3323-6_13
摘要
Droplet digital polymerase chain reaction (ddPCR) is a new quantitative PCR method based on water-oil emulsion droplet technology. ddPCR enables highly sensitive and accurate quantification of nucleic acid molecules, especially when their copy numbers are low. In ddPCR, a sample is fractionated into ~20,000 droplets, and every nanoliter-sized droplet undergoes PCR amplification of the target molecule. The fluorescence signals of droplets are then recorded by an automated droplet reader. Circular RNAs (circRNAs) are single-stranded, covalently closed RNA molecules that are ubiquitously expressed in animals and plants. CircRNAs are promising as biomarkers for cancer diagnosis and prognosis and as therapeutic targets or agents to inhibit oncogenic microRNAs or proteins (Kristensen LS, Jakobsen T, Hager H, Kjems J, Nat Rev Clin Oncol 19:188-206, 2022). In this chapter, the procedures for the quantitation of a circRNA in single pancreatic cancer cells using ddPCR are described.
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