Method and reagent for determining specific nucleotide...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C07H 21/04 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2071537

Detection of variable nucleotide(s) is based on primer extension and incorporation of detectable nucleoside triphosphates. By selecting the detection step primers from the region immediately adjacent to the variable nucleotide, this variation can be de- tected after incorporation of as few as one nucleoside triphosphate. Labelled nucleoside triphosphates matching the variable nu- cleotide are added and the incorporation of a label into the detection step primer is measured. The selection of the detection step primer is important to the method according to this invention and is dependent on the nucleotide sequence of interest. The detec- tion step primers are preferably selected so to span the region immediately toward the 3' end from the variable nucleotide to be detected. The method is useful in identifying specific point mutations and genetic variations.

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