Fluorescent multiplex amplification kit for detecting human Y-STR loci and application of fluorescence multiplex amplification kit
A Y-STR, compound amplification technology, applied in the field of molecular genetics, to achieve the effect of rapid amplification, simple operation method and good balance
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Embodiment 1
[0026] 1. Screening of 50 very low mutation Y-STR loci.
[0027] 通过对DYS618、DYS640、DYS497、DYS567、DYS590、DYS579、DYF390S1、DYS615、DYS621、DYF381S2、DYF380S1、DYS545、DYS619、DYS559、DYS595、DYS536、DYS478、DYS577、DYF405S1、DYS502、DYS642、DYS625、DYF382S1、DYS388、 DYS426、DYS435、DYS436、DYS581、DYS550、DYS453、DYS494、DYS584、DYS641、DYS477、DYS480、DYS575、DYS474、DYS588、DYS499、DYS448、DYS530、DYS538、DYS512、DYS558、DYS583、DYS434、DYS485、DYS580、DYS624、 DYS491、DYS573、DYS473、DYS571、DYS475、DYS470、DYS634、DYS455、DYS613、DYS488、DYS472、DYS489、DYS454、DYS476、DYS554、DYS561、DYS521、DYS574、DYS631、DYS525和DYS578共70个Y染色体STR基因座的 基因多态性和突变率的研究,最后筛选出DYS618、DYS640、DYS497、DYS567、DYS590、DYF390S1、DYS502、DYS615、DYS621、DYF381S2、DYF380S1、DYS545、DYS559、DYS595、DYS536、DYS478、DYS577、DYF405S1、DYS642 、DYS625、DYF382S1、DYS436、DYS550、DYS453、DYS494、DYS641、DYS477、DYS480、DYS575、DYS474、DYS588、DYS499、DYS530、DYS538、DYS512、DYS558、DYS485、DYS580、DYS624、DYS491、DYS573、DYS473、DYS571、DYS475 , DYS470, DYS634, DYS455, DYS613, DYS488, DYS472, a total of 50 very...
Embodiment 2
[0031] 1. PCR reaction buffer and amplification program adjustment
[0032] (1) The buffer of the PCR reaction (ie, A-Reaction Mix) contains various ions and dNTP mixtures necessary for the amplification reaction, and some specific PCR enhancers can also be added according to the needs of PCR. However, in order to improve the amplification specificity, amplification efficiency and anti-interference ability of the PCR reaction, it is necessary to optimize the concentration of the above components.
[0033] The optimized concentration ranges of the components contained in the PCR reaction buffer in the kit of the present invention are as follows.
[0034]
[0035] (2) The PCR amplification procedure is different from ordinary PCR in that no extension step is required. The specific amplification procedure is:
[0036]
[0037] 2. Amplification reaction system and data analysis.
[0038] The components and content of the amplification system in this kit are shown in the t...
Embodiment 3
[0041] Example 3 Different Family Male Individual Sample Test
[0042] Two male individual samples from different families were selected, and the amplification product was analyzed according to the method provided in Example 4. The results are as follows:
[0043]
[0044]
[0045] The results showed that there was only 4 steps difference between the two samples at Y37, so they might not be in the same family. The Y50 test showed that the typing of 50 loci was the same, and they belonged to the same family. Highly mutated loci may lead to false positive exclusions. The Y50 hypomutation locus could serve as strong support. It also proves that the kit has the ability to distinguish families. The kit is fully compatible with the current PCR-capillary electrophoresis platform popularized in forensic DNA laboratories. Amplify.
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