Silberklang M, Gillum A M, RajBhandary U L
Nucleic Acids Res. 1977 Dec;4(12):4091-108. doi: 10.1093/nar/4.12.4091.
A method is described for the direct sequence analysis of 20-25 nucleotides from the termini of 5'- or 3'-end-group [32P] labeled RNA. The method involves partial endonucleolytic digestion of the labeled RNA with nuclease P1 (from Penicillium citrinum) followed by separation of the partial digestion products by two-dimensional homochromatography, the nucleotide sequence being determined by mobility shift analysis. This procedure has been applied to the sequence analysis of the terminal regions of tRNAs and of high molecular weight RNA, such as messenger RNA or viral RNA. A further application involves its use in conjunction with snake venom phosphodiesterase to determine the sequence of 5'-end group labeled oligonucleotides, containing modified bases, derived from T1 or pancreatic RNase digestion of tRNA.
本文描述了一种对5'-或3'-端基[32P]标记RNA末端20 - 25个核苷酸进行直接序列分析的方法。该方法包括用核酸酶P1（来自桔青霉）对标记RNA进行部分核酸内切酶消化，然后通过二维同系层析分离部分消化产物，通过迁移率变动分析确定核苷酸序列。此程序已应用于tRNA末端区域以及高分子量RNA（如信使RNA或病毒RNA）的序列分析。进一步的应用包括将其与蛇毒磷酸二酯酶结合使用，以确定来自tRNA经T1或胰核糖核酸酶消化得到的含修饰碱基的5'-端基标记寡核苷酸的序列。
