基于离液剂的杂交溶液中寡脱氧核苷酸探针的应用。
The use of oligodeoxynucleotide probes in chaotrope-based hybridization solutions.
作者信息
Van Ness J, Chen L
机构信息
MicroProbe Corporation, Bothell, WA 98021.
出版信息
Nucleic Acids Res. 1991 Oct 11;19(19):5143-51. doi: 10.1093/nar/19.19.5143.
Abstract
Hybridization solutions containing chaotropes may be used to modulate the thermal stability (Tm or Td) of oligodeoxynucleotide (ODN) duplexes or hybrids over a 90 degrees C range. Modulation of Td allows formulation of hybridization solutions that permit ambient temperature hybridization using most combinations of probe length, probe composition, target type, and facilitates development of convenient and rapid assay formats. The conditions required to achieve ODN duplex fidelity, and optimal yields of hybridized product, are described for trichloroacetate, thiocyanate, guanidinium salts and other chaotropic salts. The effects of different solid supports on Td are described. Also, a method is presented that uses chaotropic compounds to reduce background arising from signal ODN probes in a sandwich assay hybridization format.
摘要
含有离液剂的杂交溶液可用于在90℃范围内调节寡脱氧核苷酸（ODN）双链体或杂交体的热稳定性（Tm或Td）。Td的调节使得能够配制杂交溶液，允许使用大多数探针长度、探针组成、靶标类型的组合在环境温度下进行杂交，并有助于开发方便快捷的检测形式。本文描述了实现ODN双链体保真度以及杂交产物最佳产量所需的条件，涉及三氯乙酸盐、硫氰酸盐、胍盐和其他离液盐。描述了不同固相支持物对Td的影响。此外，还提出了一种方法，该方法使用离液化合物来减少夹心测定杂交形式中信号ODN探针产生的背景。
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