Antisense interference oligonucleotide used for inhibiting primer non-specific amplification

Abstract

The invention relates to an antisense interference oligonucleotide used for inhibiting primer non-specific amplification. Through combining a primer 3' end antisense oligonucleotide, the antisense interference oligonucleotide interferes PCR primer 3' end non-specific amplification. The antisense oligonucleotide is a section of oligonucleotide with 4-14 base composed of antisense nucleotide. The basic characteristics are that the antisense base oligonucleotide maintains a Watson base pairing hybrid property, but loses the functions as PCR amplification template and primers. Antisense oligonucleotide is immobilized by using nanospheres, such that primers are indirectly immobilized to a PCR system, and hot start is sustained-released. Mineral oil is used for sealing PCR, such that aerosol cross-contamination is avoided, and PCR reaction is prevented from non-specific amplification. The antisense oligonucleotide is suitable for various PCR technologies with primer annealing and extension, including array amplification technology.

Description

Technical field: [0001] The invention belongs to the technical field of nucleic acid amplification in the field of molecular biology and molecular testing, and specifically relates to the technical field of interfering with non-specific amplification such as primer polymerization by antisense "dead" oligonucleotides competing for non-specific templates. Background technique: [0002] Nucleic acid amplification technology originated in 1971. Korana first proposed the idea of ​​in vitro nucleic acid amplification: "After DNA denaturation, hybridization with suitable primers, extension of primers with DNA polymerase, and continuous repetition of this process, tRNA genes can be cloned." The seeds of its ideas will take root and germinate when they enter the soil of the development of modern molecular biology. Finally, in 1983, Kary Mullis of the Human Genetics Laboratory of PE-Cetus Company in the United States had the inspiration of nucleic acid amplification when studying nucl...

AI Technical Summary

Problems solved by technology

Generally, dUTP is used to replace the dTTP product for aerosol recontamination of the amplified product, combined with uracil-DNA-glycosylase (UDG / UNG, US Patent 6,090,553) to selectively degrade the contaminated product. UDG / UNG is subsequently thermally denatured and inactivated by PCR, but In actual work, the degradation effect on a large number of primer-dimer-containing dUTP amplification products is limited, and it cannot eliminate or delay the Ct value of the SYBR Green I real-time fluorescent PCR background primer

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