Method for detecting variation of gene for non-diagnostic purpose based on fluorescence quenching and probe thereof
A method for detecting variation of gene based on fluorescence quenching quantification comprises: performing single base extension at a specific site of a gene to be detected with marked probes and marked dideoxyribonucleotide triphosphate; detecting fluorescence quenching values, and determining SNP of the gene to be detected. The present invention also provides an oligonucleotide probe for the method.
1. The oligonucleotide probe, comprising 15-45 nucleotides and marked with a quenching molecule (BHQ1, BHQ2, BHQ3, TAMRA, or Dabcyl or Eclipse); a distance between said quenching molecule and 3′ end of said oligonucleotide sequence marked with said fluorescence molecule (R6G, Cy3, Cy5, or FAM) is 3-35 bp, wherein said oligonucleotide probe comprises a single probe having a sequence as SEQ ID NO:3 or SEQ ID NO:6.
2. The oligonucleotide probe, comprising 15-45 nucleotides and marked with a fluorescence molecule (R6G, Cy3, Cy5, or FAM); a distance between said fluorescence molecule and 3′ end of said oligonucleotide sequence marked with said quenching molecule (BHQ1, BHQ2, BHQ3, TAMRA, or Dabcyl) is 3-35 bp, wherein said oligonucleotide probe comprises double probes respectively having sequences as SEQ ID NO:7 or SEQ ID NO:8.