MTHFR (Methylene Tetrahydrofolate Reductase) gene detection primer group, detection kit and detection method
A gene detection and primer set technology, applied in the fields of genetic engineering and molecular biology, can solve the problems of low sensitivity, high false positive rate and long period of MTHFR gene detection, improve specificity and sensitivity, reduce false positive rate, The effect of short detection period
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no. 1 example
[0030] The present invention proposes a first embodiment, a primer set for MTHFR gene detection, including at least one of the rs1801133 primer set and the rs1801131 primer set;
[0031] The rs1801133 primer set is used to amplify nucleic acid fragments containing rs1801133 sites, including rs1801133 upstream primers and rs1801133 downstream primers, and the rs1801133 upstream primers have SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 or the sequence of SEQ ID NO:7, the rs1801133 downstream primer has SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:10, SEQ the sequence of ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13 or SEQ ID NO: 14;
[0032] The rs1801131 primer set is used to amplify nucleic acid fragments containing the rs1801131 site, including rs1801131 upstream primers and rs1801131 downstream primers, and the rs1801131 upstream primers have SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 17, SEQ ID NO:18 or the sequence ...
no. 2 example
[0038] The present invention proposes a second embodiment, a MTHFR gene detection kit, containing any one of the above MTHFR gene detection primer sets.
[0039] The kit of the present invention can specifically amplify nucleic acid fragments containing rs1801133 sites and / or rs1801131 sites, reducing the possibility of amplifying non-target fragments, thereby enabling detection of rs1801133 sites and / or rs1801131 sites High sensitivity and low false positive rate.
[0040] Two sets of MTHFR gene detection primer sets are used for two rounds of nested amplification of the target site. Since there are few sequences complementary to both sets of primer sets at the same time, the possibility of non-specific amplification is reduced and the target site is increased. Sensitivity of point detection.
[0041] Preferably, the kit also includes non-fluorescent labeled oligonucleotide probes.
[0042] It should be noted that the sequence of the non-fluorescent-labeled oligonucleotide ...
no. 3 example
[0056] The present invention proposes a third embodiment, a method for MTHFR gene detection, comprising the following steps:
[0057] A. Use the rs1801133 primer set to amplify the nucleic acid fragment containing the rs1801133 site to obtain the rs1801133 amplification product; or use the rs1801131 primer set to amplify the nucleic acid fragment containing the rs1801131 site to obtain the rs1801131 amplification product;
[0058] The rs1801133 primer set is used to amplify nucleic acid fragments containing rs1801133 sites, including rs1801133 upstream primers and rs1801133 downstream primers, and the rs1801133 upstream primers have SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 or the sequence of SEQ ID NO:7, the rs1801133 downstream primer has SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:10, SEQ the sequence of ID NO: 11, SEQ ID NO: 12 or SEQ ID NO: 13;
[0059] The rs1801131 primer set is used to amplify nucleic acid fragment...
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