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Single nucleotide polymorphisms (Single Nucleotide Polymorphisms, SNPs) are the most common variations in the human genome sequence, their study helps to explain the individual phenotypic differences, different groups and individual susceptibility to complex diseases, as well as various drug tolerability and response to environmental factors. Therefore, the establishment of a rapid, accurate, high-throughput and suitable for clinical SNP genotyping method is very important. Many SNP genotyping methods have been reported, the vast majority of the need for SNP loci PCR products were purified, concentrated, which is a time-consuming and laborious work, and it is difficult to meet the future requirements of typing automation . With the rapid development of nanotechnology, nanomaterials have been gradually applied to the field of life sciences, new technologies and means for its research and development. Magnetic nanoparticles (Magnetic nanoparticles, MNPs) as an important part of the nanomaterials, because of its special physical and chemical properties and biocompatibility, has been widely used in cell separation, immunoassays, protein and enzyme immobilization and DNA testing. Enrichment of magnetic nanoparticles and bio-chip \for the detection of single nucleotide polymorphisms of methylenetetrahydrofolate reductase (MTHFR) gene 677 points (C → T), the application of this method on 126 samples of MTHFR C677T polymorphism detection. The experimental results show that, when connected downstream primer in the surface of the magnetic nanoparticles, and the template DNA in the reaction system as well as the upstream primer was amplified by PCR and the desired target fragment. When the hybridization temperature at 38 ℃, the use of fluorescent probes can accurately distinguish sample SNP genotyping, genotyping results be verified by sequencing. To further reduce genotyping cost, based on the proposed the MNPs-PCR typing method based on the use of two with a wild mutation common label (tag) probe and two-color fluorescent (Cy3, Cy5) labeled universal detecting sub, further reducing the cost of the sample of stars. This article MTHFR C677T locus of the 96 samples, and AGT gene M235T sites with this new method of high-throughput genotyping. The above-mentioned two kinds of SNP genotyping magnetic nanoparticles PCR based method not only omit the conventional method of purification, the tedious steps of the PCR product was concentrated, and to obtain a good genotyping results. Then verify that the automatic detection accuracy of the method in an automated workstation, 96 samples AGT gene M235T sites automated detection, the entire experimental operation is done automatically in 96-well PCR plate, using a common tag technology, the final a detection chip down the degeneration generic detection of fluorescently labeled quantum dots on a glass slide scan can be completed in minutes typing a large number of samples, and intuitive, accurate genotyping results is wrong with signal ratio, is a The kinds of operation is simple, fast, high-throughput, high-sensitivity typing method, a high application value.
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