Single Nucleotide Polymorphism Method Using 2-aminopurine Fluorescence Probe Recognition

The DNA molecule is the most important genetic material of human body, single base is the basic unit of the structure and function of the DNA molecule. DNA mutations and damage, in many cases, is neginning with the mutation of a single nucleotide. Therefore, to study the polymorphism of single nucleotide has a very important significance.Nowadays, the use of marked and known sequence oligonucleotide probes to identify single nucleotide polymorphisms is a important topics in the field of environmental toxicology. In recent decades, the type of fluorescent probes had increased and function developed, of which2-Aminopurine as a purine anolog is widely recognized of its own good fluorescent. In this paper, we study the changes of2-Aminopurine’s fluorescence properties in identifying single nucleotide polymorphisms process.The study consists of three main parts:(1) changing single bases in the oligonucleotide on both sides of the2-Aminopurine, formed by nucleotide ATCG, composed of16kinds of micro-environment, detecting2-Aminopurine’s fluorescent property changes in the sixteen kinds of micro-environment.(2) using2-Aminopurine probe single-strand hybridized with its complementary sequence oligonucleotides, in which contain a bases opponent to2-Aminopurine of ACGT. Detecting the fluorescence characteristics change of2-Aminopurine when bit different opponent base. Finding the relationship between the change of base type and the changes in the fluorescence properties of2-Aminopurine, which can achieve the purpose of identifying single nucleotide type.(3) using2-Aminopurine probe single-strand hybridized with its complementary sequence oligonucleotides, in its bit position with a dspacer to simulate single base deletion. Detecting the fluorescence characteristics change of 2-Aminopurine in the state where the nucleotide deletion. Finding the relationgship between2-Aminopurine’s fluorescence characteristic changes and base deletion through analysis, which can be achieved through the characterization of the fluorescence characteristics identify single nucleotide deletion behavior.We analysed the different applicabilities of2-Aminopurine in identifying single nucleotide polymorphism due to microenvironmental changes, and initially established a method on using2-Aminopurine detecting single nucleotide polymorphism in16kinds of microenvironment.