Matz M, Usman N, Shagin D, Bogdanova E, Lukyanov S
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, V-437 Moscow 117871, Russia.
Nucleic Acids Res. 1997 Jun 15;25(12):2541-2. doi: 10.1093/nar/25.12.2541.
A method for display of 3'-end restriction fragments of cDNAs is proposed, extending the idea reported recently. First, representative pools of such fragments are selectively amplified using PCR suppression effect. Then, simplified subsets of these fragments suitable for comparison by PAGE are amplified by adapter-specific primers extended by two randomly picked bases at their 3'ends. By testing all possible combinations of extended primers the whole mRNA pool may be systematically investigated. The method was applied to search for molecular regional markers of freshwater planarian Dugesia tigrina .
本文提出了一种展示cDNA 3'端限制性片段的方法,该方法拓展了近期报道的思路。首先,利用PCR抑制效应选择性扩增此类片段的代表性文库。然后,通过在3'端延伸两个随机选取碱基的接头特异性引物,扩增出适合PAGE比较的这些片段的简化亚组。通过测试延伸引物的所有可能组合,可以系统地研究整个mRNA文库。该方法已应用于寻找淡水涡虫杜氏阔盘吸虫的分子区域标记。