SNP detection experiment
SNP (SINGLE NUCLETIDE POLYMORPHISM), ie mononucleotide polymorphism Sexuality is due to a single nucleotide change. In general, only two alleles are only two alleles, so it is also called a double allele. The frequency of SNP is relatively high in the human genome, and there is a multi-state site per 1000 base. Some SNP sites will also affect the functionality of the genes, leading to biological changes or even disease. Single nucleotide polymorphism is an important basis for studying the genetic variation of human family and animal flora, therefore is widely used in group genetics research (such as the origin of biological origin, evolution and migration) and the study of disease-related genes. An important role in pharmacotics, diagnostics and biomedical research.
Service item
1, Taqman probe method u0026 EMSP; u0026 EMSP; Different SNP sites on chromosomes design PCR primers and TAQMAN probes, respectively, and perform real-time fluorescent PCR amplification. The 5 u0026 RSQUO; - End and 3 u0026 RSQUO; - Terminally marked a reporting fluorescent group and a quenching fluorescent group, respectively. When the PCR product is present in the solution, the probe is annealed with the template, i.e., a substrate suitable for the external transk activity of the nucleic acid, thereby cutting the probe 5 u0026 RSQUO; -Ted fluorescent molecules cut from the probe, destroying two PRET between the fluorescent molecules emits fluorescence. Usually used for a small amount of SNP site analysis
SnapShot method u0026 EMSP; u0026 EMSP; this technology is developed by the US Application Biological Corporation (ABI), Splitting technology based on fluorescence marked single base extension principle, also known as small sequencing, primarily for medium flux SNP classification projects. In a reaction system containing sequencing enzymes, four fluorescent labels DDNTP, close to multi-state sites 5 u0026 RSQUO;--terminal, the primer extends an base to terminate, after detecting the ABI sequencer, According to the movement position of the peak, the SNP site corresponding to the extension is determined according to the color of the peak, and the incorporated base type is obtained, thereby determining the genotype of the sample. The PCR product template can be obtained by multiple PCR reaction systems.
3, HRM Method
4, mass array method [ 123] u0026 EMSP; u0026 EMSP; MassArray molecular array technology is genetic analysis tools launched by SEQUENOM, which combines primer extension or cutting reaction with sensitive, reliable MALDI-TOF-MS technology to achieve genotyping detection. IPLEX Gold technology based on the MassArray platform can design up to 40% of PCR reactions and genotype detection, flexible experimental design, and high accuracy of the classification results. According to the application needs, when dozens to hundreds of SNP sites are tens of thousands to thousands of sample detection, MassArray has a better cost performance, especially suitable for verification of the results of all genome studies, or a limited number of research The site has been determined.
u0026 EMSP; u0026 EMSP; BEADXPRESS system using Illumina, can detect 1-384 SNP sites at the same time, often for genome chips The results are confirmed, suitable for high-throughput detection. The microbead chip has a high density, high repetitive, high sensitivity, low sample, a custom flexible, and extremely high integrated density, resulting in extremely high detection screening speed, which can significantly reduce costs in high throughput screening. 6, KASP genotyping technology KASP is an abbreviation for competitive allele specific PCR (Kompetitive Allele Specific PCR), available in a wide range of genomic DNA samples (Even some of the DNA samples of some complex genomes), accurate dual-alleles for INDELS on SNPS and specific sites. This technology is based on the specific match of the primer end base to detect IndEls (INSERTIONS AND DELETITIONS, insertion, and deletion).
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