METHOD FOR DETERMINING THE FG-2 CANOLAS GENETICITY OF A CANOLA USING PCR WITH AN END-POINT DETECTION

摘要

1. A method for determining the zygosity of a canola plant comprising the fad-2 gene, the aforementioned method comprising: obtaining a genomic DNA sample from the aforementioned canola plant; obtaining a contacted sample by contacting the aforementioned genomic DNA sample with a first primer and a second primer, wherein the aforementioned first primer preferably binds a region of the aforementioned fad-2 gene above the position of the considered single nucleotide polymorphism, the aforementioned second primer preferably binds the aforementioned fad-2 gene below the single nucleotide polymorphism in question, where the aforementioned first primer and the aforementioned second primer produce amplicon when subjected to polymerase chain reaction (PCR) conditions; subjecting the aforementioned contacted sample to PCR conditions under which the aforementioned amplicon is formed; allowing each of the aforementioned amplicon; the first fluorescent probe and the second fluorescent probe hybridize with amplicon for a predetermined period of time at temperatures from 50 to 70 ° C, wherein the aforementioned first fluorescent probe preferably hybridizes with the aforementioned amplicon when the aforementioned single nucleotide polymorphism is not present in the aforementioned amplicon, the aforementioned second fluorescent probe preferably hybridizes with the aforementioned amplicon when the aforementioned said single-nucleotide polymorphism is present; the above temperature after a period of time determined in the dispensing step