Kit and detection method for detecting EGFR gene T790M mutation
A technology of T790M-F and T790M-R is applied in the field of kits for detecting EGFR gene T790M mutation, which can solve the problems of long detection time, complicated procedures and high detection cost, and achieve the effect of reducing detection time and cost.
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Embodiment 1
[0055] This embodiment provides a kit for detecting the T790M mutation of the EGFR gene, including an isothermal amplification reaction system, a detection reaction system, and a ctDNA standard product with known T790M mutation frequency of the EGFR gene.
[0056] The isothermal amplification reaction system comprises:
[0057] The isothermal amplification reaction system includes: T790M site detection primers, Rehydration Buffer, MgA C and RPA lyophilized enzyme;
[0058] Wherein, the T790M site detection primers include primer T790M-F and primer T790M-R:
[0059] The base sequence of the primer T790M-F is as follows:
[0060] GAAATTAATACGACTCACTATAGGGCATCTGCCTCACCTCCACCGTGCAGCTCATC.
[0061] The base sequence of the primer T790M-R is as follows:
[0062] TTCCCGGACATAGTCCAGGAGGCAGCCGAAGGGC.
[0063] The detection reaction system includes: Cas13a Enzyme Mix, detection mutant probe, dNTP Mix, T7 RNAPolymerase Mix and MgCl 2 .
[0064] The detection mutant probe is T790MP...
Embodiment 2
[0082] This embodiment provides a kit for detecting the T790M mutation of the EGFR gene, including an isothermal amplification reaction system, a detection reaction system, and a ctDNA standard product with known T790M mutation frequency of the EGFR gene.
[0083] T790M site detection primer, Rehydration Buffer, MgA C and RPA lyophilized enzyme;
[0084] Wherein, the T790M site detection primers include primer T790M-F and primer T790M-R:
[0085] The base sequence of the primer T790M-F is as follows:
[0086] GAAATTAATACGACTCACTATAGGGCATCTGCCTCACCTCCACCGTGCAGCTCATC.
[0087] The base sequence of the primer T790M-R is as follows:
[0088] TTCCCGGACATAGTCCAGGAGGCAGCCGAAGGGC.
[0089] The detection reaction system includes: Cas13a Enzyme Mix, detection mutant probe, dNTP Mix, T7 RNAPolymerase Mix and MgCl 2 .
[0090] The detection mutant probe is T790MProbe, and the base sequence of the T790MProbe is:
[0091] TAATACGACTCACTATAGGGGATTTAGACTACCCCAAAAACGAAGGGGACTAAAACGCATCA...
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