Kit for detecting gastric cancer risk gene and using method thereof
A gene detection and kit technology, which is applied in the field of gastric cancer risk gene detection kits, can solve the problems of increasing the risk of gastric cancer and achieve the effects of convenient and accurate application, pollution prevention and simple operation
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Embodiment 1
[0040] Such as figure 1 As shown, a gastric cancer risk gene detection kit of the present invention includes detection reagents for detecting one or more of 11 single nucleotide polymorphism sites in 9 genes, 11 of 9 genes The single nucleotide polymorphism sites are specifically: T→C of COX-2 gene rs5273 and G→C variation of rs20417, A→G variation of IL-10 gene rs1800896; C→G variation of P53 gene rs1042522; T→C of PARP-1 gene rs1136410 and A→G variation of rs3219145; A→G variation of PLCE1 gene rs2274223; T→C variation of RKAA1 gene rs13361707; C→T variation of PSCA gene rs2294008; G→G variation of SOD1 gene rs4998557 A variation; T→C variation of SOD2 gene rs4880.
[0041] The detection reagents include: PCR reagents and genotype determination reagents.
[0042] Wherein, the genotype determination reagents are Taqman probe fluorescent PCR reagents, Snapshot reagents, reagents for fluorescent PCR method, reagents for constant temperature amplification-fluorescence method, ...
Embodiment 2
[0108] The difference between this example and Example 1 lies in that the methods for analyzing and determining the genotype of the target sequence are different.
[0109] 3. Analysis and determination of target sequence genotype
[0110] Generation Sequencing
[0111] Nucleotide sequence determination:
[0112] 1. Glue recovery:
[0113] 1.1 Add Binding Buffer 30ul to the tube containing gel recovery and incubate at 55°C for 7min until it completely dissolves;
[0114] 1.2 After the gel is melted, take out the column tube, mark it, transfer the liquid into the column tube, centrifuge at 10000rpm / 1min, pour off the supernatant after completion, add 700ulSpw, let stand for 2-3min, centrifuge at 10000rpm / 1min, pour Drop the supernatant;
[0115] 1.3 Repeat the above 1.2 operation once;
[0116] 1.4 After pouring off the supernatant, the column tube is idling at 1000rpm / 1min;
[0117] 1.5 Add 15ulElution Buffer to the center of the column, let it stand for 2-3min, and centr...
Embodiment 3
[0130] The difference between this example and Example 1 lies in that the methods for analyzing and determining the genotype of the target sequence are different.
[0131] 3. Analysis and determination of target sequence genotype
[0132] Taqman probe fluorescent PCR method
[0133] reaction system:
[0134] Component
Volume
Final Concentration
1pg-1ug
Forward primer (10uM)
0.4ul
0.2uM
Reverse primer (10uM)
0.4ul
0.2uM
Probe(10uM)
0.4ul
0.2uM
qPCR SuperMix
10ul
1×
Passive reference Dye (50x)
0.4ul
1×
Variable
-
Total volume
20ul
-
[0135] Reaction conditions:
[0136]
[0137] Amplification primers and probes:
[0138]
[0139]
[0140] In the present invention, other methods can also be used for the analysis and determination of the target sequence genotype. Such as:
[0141] 1. Chip detection based...
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