Thermodynamic optimization-based high specificity nucleic acid hybridization method

A thermodynamically optimized and highly specific technology, applied in the fields of biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as difficult to fully satisfy

Active Publication Date: 2015-04-15
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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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 misma

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  • Thermodynamic optimization-based high specificity nucleic acid hybridization method
  • Thermodynamic optimization-based high specificity nucleic acid hybridization method
  • Thermodynamic optimization-based high specificity nucleic acid hybridization method

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Embodiment 1

[0072] (1) Design and preparation of probes

[0073] First, strand displacement probes were designed according to the target microRNA-125 to be detected. The specific design method is to design a nucleic acid single strand that is completely complementary to the target of interest, and then add several bases to one end of the nucleic acid single strand, called the complementary strand. When designing the other strand of the probe, it should be identical to the partial sequence of the target sequence to be detected, which is called the release strand. Adjust the base sequence of certain positions in the probe so that the standard enthalpy of reaction Δ r h 0 and reaction standard entropy Δ r S 0 The value is approximately equal to 0. The designed probes will be synthesized by the company. After the probe sequence is synthesized, the complementary strand and the release strand are mixed and hybridized according to a certain ratio and diluted to an appropriate concentration...

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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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Application Information

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IPC IPC(8): C12Q1/68
Inventor 毛红菊武振华白亚楠金庆辉赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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