Application of SNP loci of Nodal, GDF1 genes
A site and gene technology, applied in the field of Nodal gene and GDF1 gene SNP site and related kits, can solve the problems of unconfirmed, decreased enzyme activity, unidentified, etc., and achieve the effect of good sensitivity
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Embodiment 1
[0046] Embodiment 1, Nodal gene and CHD
[0047] 1.1 Extraction of genomic DNA (refer to kit instructions)
[0048] 1.2 Primer design and synthesis
[0049] SNP sites rs1904589 and rs10999334, rs2279253 and rs2279254 of Nodal gene were selected for PCR-DHPLC detection. The flanking sequences were retrieved through the U.S. National Library of Medicine Pubmed engine (http: / / www.ncbi.nlm.nih.gov / sites / snp), we used Premier Primer5.0 combined with Premier3.0 to design primers online, and DNAMAN6 .0.40 software testing is qualified, and then use the WAVEMAKE Software Version4.1 software to carry out DHPLC simulation detection on the fragments to be amplified with different primers, and select the most suitable primer sequence for on-board detection. The Navigator software package of the system can be used for DHPLC detection temperature Make predictions. If there is a significant difference between the chromatographic elution peaks of the test sample and the known wild-type con...
Embodiment 2
[0134] Embodiment 2, GDF1 gene and CHD
[0135] 2.1 Extraction of genomic DNA (refer to kit instructions)
[0136] 2.2 method (embodiment 1)
[0137] 2.2.1 Primer design and detection of three polymorphic sites (rs7025622, rs4808870r, s2075762)
[0138] We also used Premier Primer5.0 combined with Premier3.0 to design primers online, and the DNAMAN6.0.40 software tested the primers to be qualified, and then used the WAVEMAKE Software Version4.1 software to perform DHPLC simulation detection on the fragments to be amplified with different primers, and selected the best Primer sequences suitable for on-board detection, and use the system's Navigator software package to predict the detection temperature of DHPLC. The determined primers (Table 2-1) were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. and purified by HAP.
[0139] Table 2-1 PCR amplification primers, product length, annealing and DHPLC temperature of the three SNPs of GDF1
[0140] ...
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