Architecture et Réactivité de l'ARN - CNRS UPR 9002, Institut de Biologie Moléculaire et Cellulaire, Université de Strasbourg, Strasbourg, France.
Plateforme protéomique Strasbourg Esplanade FRC1589 du CNRS, Strasbourg, France.
Methods Mol Biol. 2020;2113:101-110. doi: 10.1007/978-1-0716-0278-2_8.
RNA modification mapping by mass spectrometry (MS) is based on the use of specific ribonucleases (RNases) that generate short oligonucleotide digestion products which are further separated by nano-liquid chromatography and analyzed by MS and MS/MS. Recent developments in MS instrumentation allow the possibility to deeply explore posttranscriptional modifications. Notably, development of nano-liquid chromatography and nano-electrospray drastically increases the detection sensitivity and allows the identification and sequencing of RNA digested fragments separated and extracted from two-dimensional polyacrylamide gels, as long as the mapping and characterization of ribonucleotide modifications.
基于质谱（MS）的 RNA 修饰图谱分析是基于使用特定的核糖核酸酶（RNases），这些酶会产生短的寡核苷酸消化产物，然后通过纳升液相色谱进一步分离，并通过 MS 和 MS/MS 进行分析。MS 仪器的最新发展使得深入探索转录后修饰成为可能。值得注意的是，纳升液相色谱和纳升电喷雾的发展极大地提高了检测灵敏度，并允许从二维聚丙烯酰胺凝胶中分离和提取的 RNA 消化片段进行鉴定和测序，只要对核糖核苷酸修饰进行定位和特征分析。
