Dickson E, Pape L K, Robertson H D
Nucleic Acids Res. 1979 Jan;6(1):91-110. doi: 10.1093/nar/6.1.91.
A method is described for the initial steps of sequence analysis of RNase T1-and pancreatic RN-ase-resistant oligonucleotides of RNA containing cytidylate residues labeled in vitro with 125I. In many cases an oligonucleotide sequence can be deduced from a consideration of (i) its relative position in the two-dimensional fingerprint (with DEAE thin layer homochromatographic second dimension), (ii) its electrophoretic mobility on DEAE paper at pH 1.9, and (iii) identification of its products of further enzymatic digestion by comparison with a set of marker oligonucleotides. Additional methods including analysis of oligonucleotides following chemical blocking of uridylate residues with CMCT and analysis of products of incomplete enzymatic digestion are also discussed.
本文描述了一种对含有经体外125I标记的胞苷酸残基的RNA的核糖核酸酶T1和胰核糖核酸酶抗性寡核苷酸进行序列分析初始步骤的方法。在许多情况下,可通过考虑以下因素推导出寡核苷酸序列:(i) 其在二维指纹图谱(采用DEAE薄层层析同色层析第二维)中的相对位置;(ii) 其在pH 1.9条件下在DEAE纸上的电泳迁移率;(iii) 通过与一组标记寡核苷酸比较鉴定其进一步酶切产物。还讨论了其他方法,包括用CMCT对尿苷酸残基进行化学封闭后对寡核苷酸的分析以及不完全酶切产物的分析。