Rychlik W, Rhoads R E
Department of Biochemistry, University of Kentucky, Lexington 40536.
Nucleic Acids Res. 1989 Nov 11;17(21):8543-51. doi: 10.1093/nar/17.21.8543.
A method is presented for choosing optimal oligodeoxyribonucleotides as probes for filter hybridization, primers for sequencing, or primers for DNA amplification. Three main factors that determine the quality of a probe are considered: stability of the duplex formed between the probe and target nucleic acid, specificity of the probe for the intended target sequence, and self-complementarity. DNA duplex stability calculations are based on the nearest-neighbor thermodynamic values determined by Breslauer et al. [Proc. Natl. Acad. Sci. U.S.A. (1986), 83: 3746]. Temperatures of duplex dissociation predicted by the method described here were within 0.4 degrees C of the values obtained experimentally for ten oligonucleotides. Calculations for specificity of the probe and its self-complementarity are based on a simple dynamic algorithm.
本文介绍了一种选择最佳寡脱氧核糖核苷酸的方法，这些寡核苷酸可作为滤膜杂交的探针、测序引物或DNA扩增引物。考虑了决定探针质量的三个主要因素：探针与靶核酸形成的双链体的稳定性、探针针对预期靶序列的特异性以及自身互补性。DNA双链体稳定性计算基于Breslauer等人确定的最近邻热力学值[《美国国家科学院院刊》(1986年)，83: 3746]。此处所述方法预测的双链体解离温度与十种寡核苷酸实验获得的值相差在0.4摄氏度以内。探针特异性及其自身互补性的计算基于一种简单的动态算法。
