使用 mA-RT-QPCR 进行残留特异性 mA RNA 甲基化的相对定量。

Relative Quantification of Residue-Specific mA RNA Methylation Using mA-RT-QPCR.

机构信息

Department of Genetics, Physical Anthropology and Animal Physiology, University of the Basque Country (UPV-EHU), Leioa, Spain.

BioCruces Bizkaia Health Research Institute, Barakaldo, Spain.

出版信息

Methods Mol Biol. 2021;2298:185-195. doi: 10.1007/978-1-0716-1374-0_12.

DOI:10.1007/978-1-0716-1374-0_12

PMID:34085246

Abstract

Technological advances in high-throughput sequencing in combination with antibody enrichment and/or induced nucleotide-specific chemical modifications have accelerated the mapping of epitranscriptomic modifications. However, site-specific detection and quantification of mA are still technically challenging. Here, we describe a simple RT-QPCR-based approach for the relative quantification of candidate mA regions that takes advantage of the diminished capacity of BstI enzyme to retrotranscribe mA residues.

摘要

高通量测序技术的进步，结合抗体富集和/或诱导的核苷酸特异性化学修饰，加速了对表转录组修饰的定位。然而，mA 的特异性检测和定量仍然具有技术挑战性。在这里，我们描述了一种基于简单 RT-QPCR 的相对定量候选 mA 区域的方法，该方法利用了 BstI 酶逆转录 mA 残基的能力下降的优势。