Method for rapidly detecting mutation of KRAS gene
A rapid and genomic technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of inaccurate detection and low sensitivity, and achieve the effect of simple result analysis, high sensitivity, and optimized amplification effect
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[0032] The present invention combines molecular beacon probes with extension-blocking primers, and utilizes a polymerase without 5'→3' exonuclease activity to rapidly detect KRAS mutations in vitro, which not only saves consumables, but also has high sensitivity and high specificity The characteristics make the test results more reliable.
[0033] Extension-Refractory primers (Extension-Refractory primers) are primers that can form a "neck loop" structure with their own extension products; in addition to the normal primer sequence, the 5' end of the extension-refractory primer contains an additional base sequence, This sequence can be complementary to a base sequence including the mutation site in the single strand of the product obtained by its own extension, forming a "neck loop" structure; due to the difference in sequence between the wild-type and mutant templates, the formed "neck loop" There is a significant difference in the strength (Tm value) of the "ring" structure. ...
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