Zatsepin Timofei S, Varizhuk Anna M, Dedkov Vladimir G, Shipulin German A, Aralov Andrey V
Skolkovo Institute of Science and Technology, Skolkovo, Moscow, Russia.
Department of Chemistry, Lomonosov Moscow State University, Moscow, Russia.
Methods Mol Biol. 2019;1973:281-297. doi: 10.1007/978-1-4939-9216-4_18.
We developed a new technique suitable for improved detection of low-copy dsRNA using modified oligonucleotides as primers in RT-qPCR. Insertion of G-clamp residues into primers significantly improves thermal stability of duplexes with RNA without decrease of hybridization selectivity. The applicability of modified primers is demonstrated for detection of low-copy Kemerovo virus dsRNA.
我们开发了一种新技术，适用于在RT-qPCR中使用修饰的寡核苷酸作为引物来改进对低拷贝双链RNA的检测。在引物中插入G-钳位残基可显著提高与RNA双链体的热稳定性，而不会降低杂交选择性。修饰引物的适用性在检测低拷贝克麦罗沃病毒双链RNA中得到了证明。
