Method for amplifying and sequencing nucleic acid based on solid phase carrier

Abstract

The invention relates to the field of gene engineering, and provides a method for amplifying and sequencing nucleic acid based on a solid phase carrier. At least one template sequence is fixed at the surface of the solid phase carrier, and the method comprises the following steps: A, combining a primer with a primer binding site on the surface of the solid phase carrier to obtain an amplifying carrier fixed with the primer; and B, amplifying the template sequence on the amplifying carrier to obtain a amplifying product. The method is based on the solid phase carrier fixed with the template sequence on the surface to amplify, can fix the amplifying product on the surface of the solid phase carrier, and improves the combining amount of the amplifying product on the surface of the solid phase carrier; and in addition, the solid phase carrier obtained by the amplifying method is utilized to sequence, so that a detection signal of the sequencing is strengthened, the requirements on a detection instrument are reduced, and the accuracy of the sequencing is improved in equal detection conditions.

Description

Technical field [0001] The present invention relates to the field of genetic engineering, and more specifically, to a method for amplification and nucleic acid sequencing based on a solid-phase carrier. Background technique [0002] Polymerase Chain Reaction (PCR) is a method of enzymatically synthesizing and amplifying DNA fragments in vitro. It consists of high temperature denaturation, low temperature annealing, and suitable temperature extension reactions that form a cycle, which is carried out in cycles to expand the target DNA. increase. After the emergence of PCR technology, it has been continuously improved according to different needs and has been widely used in the fields of gene isolation, cloning and nucleic acid sequence detection. [0003] The prior art provides a method for amplification based on a solid-phase carrier, and its realization steps include: (1) mixing a sample to be tested with a solid-phase carrier with hydrophilic positive ions so that the sample to b...

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Problems solved by technology

[0004] Using this prior art to amplify requires a large number of nucleic acid molecules as templates. When there are fewer templates on the surface of the solid-phase carrier, the smooth progress of the amplification cannot be guaranteed, thus reducing the utilization rate of the solid-phase carrier and increasing the cost; Sequencing with the solid-phase carrier after the amplification reaction is completed, it is easy to appear that the target nucleic acid sequence cannot be accurately read due to the fact that the surface of the solid-phase carrier has too little target nucleic acid and the detection signal is too weak. High requirements for detection instruments and limitations in the accuracy of sequencing

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