Same sequence primer transpositional nucleic acid amplification technology

Abstract

The invention provides a same sequence primer transpositional nucleic acid amplification technology, which comprises the steps of: (1) linking the artificial sequence to the upstream of the target specific primer 5', and forming a chimeric primer; (2) taking the chimeric primer as an amplification target template to produce a secondary template with the two ends having artificial sequences; and (3) taking the artificial sequences with the same sequence of 65%-75% as the transpositional primer amplification secondary template. The technology is applicable to clinic molecular diagnosis, basically can utilize the primer design to prevent the false positive result from generating, and is particularly applicable to communicable disease nucleic acid detection and accurate diagnosis. The technology is particularly suitable to multiple PRC amplification technology, is nearly designed by aiming at the multiple PRC amplification technology, and the redundant and extraordinarily complicated primers of the multiple PRC amplification technology can only adopt the primer transpositional design to eliminate the nonspecific amplification of the primer polymer.

Description

Technical field: [0001] The invention belongs to the technical field of nucleic acid amplification in the fields of molecular biology and molecular diagnosis, and in particular relates to a nucleic acid amplification technology using a pair of primers with most of the same intermediate sequence in order to reduce the polymerization non-specific amplification between primers. Background technique: [0002] Nucleic acid amplification technology originated in 1971. Korana proposed the concept 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, the tRNA gene can be cloned." It also developed and matured together with the progress of modern molecular biology. 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 nucleic acid sequencing methods:...

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

Images