Asseline Ulysse, Chassignol Marcel, Aubert Yves, Roig Victoria
Centre de Biophysique Moléculaire CNRS. UPR 4301, affiliated with the University of Orléans and with INSERM, Rue Charles Sadron, 45071, Orléans Cedex 02, France.
Org Biomol Chem. 2006 May 21;4(10):1949-57. doi: 10.1039/b602262f. Epub 2006 Mar 31.
This paper describes the design of terminal-mismatch discriminating fluorescent oligonucleotides (TMDFOs). The method is based on the use of sets of oligo-2'-deoxyribonucleotide probes linked via their 5'-ends, and varying-sized flexible polymethylene chains, to thiazole orange, with the linker being attached to the benzothiazole moiety. The sequence of each set of labelled probes was identical and complementary to the sequence to be analyzed on the single-stranded nucleic acid target except at the interrogation position, located at the 5'-end of the probes in a position adjacent to the attachment site of the label, where each of the four nucleic bases were incorporated. This work allowed the selection of probes showing, upon their hybridization with the target sequence, good discrimination between the matched and the mismatched duplexes under non-stringent conditions, with the mismatched duplexes being more fluorescent than the perfectly matched ones.
本文描述了末端错配鉴别荧光寡核苷酸（TMDFOs）的设计。该方法基于使用通过其5'端连接的一组寡聚2'-脱氧核糖核苷酸探针，以及不同长度的柔性聚亚甲基链，连接到噻唑橙上，连接子连接到苯并噻唑部分。每组标记探针的序列相同，并且与单链核酸靶标上要分析的序列互补，除了在位于探针5'端且与标记附着位点相邻的询问位置处，该位置掺入了四种核酸碱基中的每一种。这项工作使得能够选择在与靶序列杂交时，在非严格条件下对匹配和错配双链体具有良好鉴别的探针，错配双链体比完全匹配的双链体更具荧光性。
