Verlaan-de Vries M, Bogaard M E, van den Elst H, van Boom J H, van der Eb A J, Bos J L
Gene. 1986;50(1-3):313-20. doi: 10.1016/0378-1119(86)90335-5.
To analyze human tumors for the presence of mutated ras oncogenes, a procedure was developed based on selective hybridization of mutation-specific oligodeoxynucleotide probes to genomic DNA [Bos et al., Nucl. Acids Res. 12 (1984) 9155-9163]. We have improved this procedure both in sensitivity and speed by including an in vitro amplification step of ras-specific sequences. This amplification step has first been described by Saiki et al. [Science 230 (1985) 1350-1353] and results in a more than 10(4)-fold increase in the sequence which might contain the mutation. Furthermore, we have improved the selectivity of our hybridizations. As a result, mutated ras oncogenes can now be detected with a dot-blot screening procedure requiring less than 1 microgram of tumor DNA.
为了分析人类肿瘤中是否存在突变的ras癌基因，我们开发了一种基于突变特异性寡脱氧核苷酸探针与基因组DNA选择性杂交的方法[Bos等人，《核酸研究》12 (1984) 9155 - 9163]。我们通过加入ras特异性序列的体外扩增步骤，在灵敏度和速度方面改进了该方法。这一扩增步骤首先由Saiki等人描述[《科学》230 (1985) 1350 - 1353]，它使可能包含突变的序列增加了10⁴倍以上。此外，我们还提高了杂交的选择性。结果，现在可以用斑点印迹筛选法检测突变的ras癌基因，该方法所需的肿瘤DNA不到1微克。
