Probe, primer and kit for detecting talent gene
A kit and gene technology, applied in the field of molecular biology, can solve the problems of cumbersome detection steps, low sensitivity and poor specificity, and achieve the effects of fast detection speed, high detection sensitivity and simple operation.
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Embodiment 1
[0037]1. Detection of the present invention with genetically engineered recombinant plasmids, with the rs4680 mutant as the detection object. Plasmids with rs4680 mutation gene constructed by genetic engineering were used as detection objects. Plasmid pCOMTM was a plasmid with rs4680 mutation; plasmid pCOMTW was a plasmid with wild-type rs4680 gene. These plasmids were dissolved in TE buffer (10mM Tris-HCl, 1mMEDTA, PH=8.0), the mass and concentration were measured by UV spectrophotometer, the plasmid copy number was calculated, and 10000, 1000, 100, 10 copies were obtained by gradient dilution with TE buffer / ul of pCOMTM and pCOMTW plasmid solutions, and then used as templates for PCR reactions.
[0038] 2. Synthesis of COMT-specific primers and probes: According to the mutation site of the human COMT gene (rs4680), design 2 probes, 5' end labeled FAM (green fluorescence) or VIC (yellow fluorescence), 3' end labeled MGB The quenching group was synthesized and labeled by Sha...
Embodiment 2
[0063] 1. The present invention is detected by genetic engineering recombinant plasmids, and the rs1456031 mutant is used as the detection object. Plasmids with rs1456031 mutation gene constructed by genetic engineering were used as detection objects. Plasmid pFOXP2M was a plasmid with rs1456031 mutation; plasmid pFOXP2W was a plasmid with wild-type rs1456031 gene. These plasmids were dissolved in TE buffer (10mM Tris-HCl, 1mMEDTA, PH=8.0), the mass and concentration were measured by UV spectrophotometer, the plasmid copy number was calculated, and 10000, 1000, 100, 10 copies were obtained by gradient dilution with TE buffer / ul of the pFOXP2M and pFOXP2W plasmid solutions were then used as templates for PCR reactions.
[0064] 2. Synthesis of FOXP2-specific primers and probes: According to the mutation site of human FOXP2 gene (rs1456031), design 2 probes, 5' end labeled with FAM (green fluorescence) or VIC (yellow fluorescence), 3' end labeled with MGB The quenching group w...
Embodiment 3
[0089] 1, detect the sensitivity of the present invention with genetic engineering recombination plasmid, the plasmid with COMT mutation gene constructed through genetic engineering is as detection object, and plasmid pCOMTM is the plasmid with RS4680 mutation; Plasmid pCOMTW is the plasmid with wild-type COMT gene . These plasmids were dissolved in TE buffer (10mMTris-HCl, 1mMEDTA, PH=8.0), the mass and concentration were measured by UV spectrophotometer, the plasmid copy number was calculated, and 100 copies / ul pCOMTM and pCOMTW plasmids were obtained by diluting with TE buffer solution, which then served as a template for the PCR reaction.
[0090] 2. Synthesis of COMT-specific primers and probes: According to the mutation site of the human COMT gene, design 2 probes, the 5' end is labeled with FAM (green fluorescence) or VIC (yellow fluorescence), and the 3' end is labeled with MGB quencher group; synthesized and labeled by Shanghai Shining Molecular Biotechnology Co., Lt...
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