One-tube kit for detecting KRAS and BRAF gene mutations in plasma free DNA
A kit and gene technology, applied in the field of molecular biology, to achieve the effects of saving material and labor costs, improving throughput, and quickly and clearly distinguishing results
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Embodiment 1
[0095] 1. KRAS gene mutation detection
[0096] Take the detection of KRAS gene GGT>GCT (G12A) mutation in clinically collected plasma samples as an example.
[0097] KRAS-G12S-F:-ATATAAACTTGTGGTAGTTGGAGCAA- (SEQ ID NO: 1)
[0098] KRAS-G12R-F:-AATATAAACTTGTGGTAGTTGGAACTC- (SEQ ID NO: 2)
[0099] KRAS-G12C-F:-ATATAAACTTGTGGTAGTTGGTGTTT- (SEQ ID NO: 3)
[0100] KRAS-G12S-R:-ACCTCTATTGTTGGATCATATTCG- (SEQ ID NO:4)
[0101] KRAS-G12D-R:-GCACTCTTGCCTACGCCTT- (SEQ ID NO: 6)
[0102] KRAS-G12A-R:-GGCACTCTTGCCTACGTCAG- (SEQ ID NO: 7)
[0103] KRAS-G12V-R:-GCACTCTTTGCCTACGCGAA- (SEQ ID NO:8)
[0104] KRAS-G12D-F:-CACATTTTCATTATTTTATTATAAGGC- (SEQ ID NO:9)
[0105] KRAS-Probe1:FAM-CCTTGACGATACAGCTAATTCAGAATCATTT-BHQ1 (SEQ ID NO:5)
[0106] KRAS-Probe2:FAM-CACAAGTTTATATTCAGTCATTTTCAGCAGGC-BHQ1 (SEQ ID NO: 10)
[0107] KRAS-Blocker1:GCCTAC G CCACCA G CTCCAACT-Pho (underlined bases are marked for nucleic acid modification, 3' -pho indicates phosphorylation modification) (SEQ ID ...
Embodiment 2
[0132] 1. BRAF gene mutation detection
[0133] Take the detection of BRAF gene V600E mutation in clinically collected plasma samples as an example.
[0134] KRAS-G12S-F:-ATATAAACTTGTGGTAGTTGGAGCAA- (SEQ ID NO: 1)
[0135] KRAS-G12R-F:-AATATAAACTTGTGGTAGTTGGAACTC- (SEQ ID NO: 2)
[0136] KRAS-G12C-F:-ATATAAACTTGTGGTAGTTGGTGTTT- (SEQ ID NO: 3)
[0137] KRAS-G12S-R:-ACCTCTATTGTTGGATCATATTCG- (SEQ ID NO:4)
[0138] KRAS-G12D-R:-GCACTCTTGCCTACGCCTT- (SEQ ID NO: 6)
[0139] KRAS-G12A-R:-GGCACTCTTGCCTACGTCAG- (SEQ ID NO: 7)
[0140] KRAS-G12V-R:-GCACTCTTTGCCTACGCGAA- (SEQ ID NO:8)
[0141] KRAS-G12D-F:-CACATTTTCATTATTTTATTATAAGGC- (SEQ ID NO:9)
[0142] KRAS-Probe1:FAM-CCTTGACGATACAGCTAATTCAGAATCATTT-BHQ1 (SEQ ID NO:5)
[0143] KRAS-Probe2:FAM-CACAAGTTTATATTCAGTCATTTTCAGCAGGC-BHQ1 (SEQ ID NO: 10)
[0144] KRAS-Blocker1:GCCTAC G CCACCA G CTCCAACT-Pho (underlined bases are marked for nucleic acid modification, 3' -pho indicates phosphorylation modification) (SEQ ID NO: 11) ...
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