Kit for simultaneously detecting human MTHFR and MTRR genes
A kit and technology for human detection, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of inability to realize automation and incomplete detection.
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Embodiment 1
[0080] Embodiment 1: the composition of kit
[0081] The composition of the kit of the present invention is shown in Table 1:
[0082] Table 1
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Embodiment 2
[0086] Example 2: DNA detection
[0087] 1. Whole blood pretreatment: use the whole blood sample processing reagent independently developed by Zhengzhou Antu Bioengineering Co., Ltd. to process the whole blood sample, and the specific operation is carried out according to the instructions.
[0088] 2. Nucleic acid extraction: Use nucleic acid extraction or purification reagents independently developed by Zhengzhou Antu Bioengineering Co., Ltd., and follow the instructions for specific operations.
[0089] 3. Build a reaction system
[0090] Calculate the number of samples required, and increase the amount of reagents in proportion to the amount in the table below, and only make one copy of negative and positive quality controls.
[0091] Table 2
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[0093] 4. On-board testing
[0094] Directly use the fluorescent quantitative PCR instrument for detection, the amplification program is set to 50°C for 2min, 60°C for 15min, 95°C for 1min, (95°C for 5s,
[0095] 60°C...
Embodiment 3
[0100] Embodiment 3: the main performance index of this kit
[0101] 66, 677, and 1298 wild, mutant, and heterozygous whole blood clinical samples were screened by sequencing, purified using whole blood processing reagents and nucleic acid extraction and purification reagents independently developed by Zhengzhou Antu Bioengineering Co., Ltd., and diluted with normal saline to 1ng / μL, using this kit for detection, the results are shown in Tables 4-5, where "no Ct" means that there is still no amplification signal value after 45 cycles of amplification.
[0102] Table 4
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[0105] table 5
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[0108] Table 6
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[0111] It can be seen from Tables 4 to 6 that the detection results of known samples show that the detection results of the kit provided by the present invention are completely consistent with the gold standard sequencing results, and the accuracy rate can reach 100%.
[0112] Table 7
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