Multi-methylation specific PCR primer design method and system

Abstract

The invention discloses a multi-methylation specific PCR primer design method and system. The method comprises the steps: setting parameters according to user requirements, designing and screening primers for a target methylation site, pairing the screened multiple methylation site primer pairs in pairs, checking the compatibility, selecting a maximum number of compatible multiple primer combinations, evaluating the designed multiple primer combinations, and determining whether the primers need to be redesigned. According to the invention, multi-methylation specific PCR primer design of a super-long sequence can be realized, secondary structures such as dimer / hairpin and the like between the interiors of a single pair of primers and between multiple pairs of primers can be effectively reduced, and non-specific amplification in a genome can be effectively reduced, the designed primers are high in specificity, the operability and accuracy of the multiplex methylation specific PCR experiment testing process are improved, and the working efficiency is greatly improved.

Description

technical field [0001] The invention relates to the field of DNA methylation detection, in particular to a method and system for designing multiple methylation-specific PCR primers. Background technique [0002] The relationship between human phenotypes and human genome mutations has gradually become clear since the development of research such as the Human Genome Project. About 88 million mutations (84.7 million single nucleotide mutations, 3.6 million short insertions or deletions) and 6 structural variants have been identified in 2015. 2001 Online Database of Human Medical Genetics (OMIM TM ) has 13,005 entries related to research results on human genome mutations and human diseases. Human genome mutations have been gradually applied in human life, such as drug guidance, drug resistance detection, prenatal detection, and tumor detection. [0003] Epigenetics is a branch of genetics that studies the heritable changes in gene expression without changing the nucleotide se...

AI Technical Summary

Problems solved by technology

However, due to the conversion of unmethylated C to T after genomic DNA is converted by sulfite, the complexity of the genome is reduced, resulting in reduced primer complexity, increased primer-dimers, secondary structures, and non-specific amplification, further increasing the Difficulty in designing multiple methylation-specific PCR primers, and conventional common multiplex PCR primer design software cannot be applied to the design of multiple methylation-specific PCR primers

At present, there is a lack of a method that can design multiple PCR primers for multiple methylation sites, and ensure that there is no secondary structure such as dimer / hairpin between a single pair of primers and between multiple pairs of primers, and it can be specifically amplified in the genome. Added systems and methods

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