Thermodynamic optimization-based high specificity nucleic acid hybridization method

Abstract

The invention relates to a thermodynamic optimization-based high specificity nucleic acid hybridization method which is as follows: the method is mainly based on a nucleic acid reaction thermodynamic nearest neighbor model, DNA and RNA reaction thermodynamic parameters are calculated, the theoretical limit of the probe specificity can be theoretically illuminated, the specific differences and thermodynamic properties of different single base mutation can be calculated simultaneously, different nucleic acid reaction results can be simulated and predicted on a computer; and a theoretically optimized high specificity nucleic acid probe can be designed. The specificity of the probe designed according to thermodynamic calculation theoretically achieves optimal results; stability is good, the probe can be stored for a long time; the detection process is particularly simple; the detection time is short; the detection process is not influenced by ambient temperature and not influenced by solution salt ion concentration, and the thermodynamic optimization-based high specificity nucleic acid hybridization method can meet the clinical demand, and has a good application prospect in medical aspects.

Description

technical field [0001] The invention belongs to the field of nucleic acid hybridization, in particular to a highly specific nucleic acid hybridization method based on thermodynamic optimization. Background technique [0002] Nucleic acid is the basic component of all biological cells, encoding and regulating the expression of genetic information in organisms. It plays an important role in the growth, development, reproduction, inheritance and variation of organisms and other major life phenomena. The biological importance of nucleic acid has led to the wide application of nucleic acid probes and primers in biotechnology such as: polymerase chain reaction (PCR), biochip (microarray) and fluorescence in situ hybridization. [0003] A key attribute of nucleic acids for use in biology, biotechnology, and nanobiotechnology is their predictable and specific Watson–Crick hybridization complementarity. However, unless near the melting temperature, the thermodynamic gain of many co...

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Problems solved by technology

[0005] Generally speaking, an ideal nucleic acid probe should meet three conditions: high affinity (capable of firmly binding to complementary targets), high selectivity (not binding to mismatched sequences), and high stability (environmental factors are not easily affected, different laboratory test results are consistent), which is difficult to fully satisfy in actual practice

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