Electrochemical Polynucleotide Detection Comprising Ligation

Abstract

Disclosed, for example, are methods comprising cleaving an uncleaved probe to form a cleaved oligonucleotide flap, forming a hybridization complex between the cleaved oligo-nucleotide flap, a bridging oligonucleotide, and a capture oligonucleotide that is immobilized on a surface, such that the oligonucleotide flap and the capture oligonucleotide are hybridized to immediately adjacent, complementary regions of the bridging oligonucleotide, ligating the oligonucleotide flap to the capture oligonucleotide to form an immobilized ligation product, and detecting the ligation product.

1 . A method comprising
 cleaving an uncleaved probe to form a cleaved oligonucleotide flap,   forming a hybridization complex between the cleaved oligonucleotide flap, a bridging oligonucleotide, and a capture oligonucleotide that is immobilized on a surface, such that the oligonucleotide flap and the capture oligonucleotide are hybridized to immediately adjacent, complementary regions of the bridging oligonucleotide,   ligating the oligonucleotide flap to the capture oligonucleotide to form an immobilized ligation product, and   detecting the ligation product.   
     
     
         2 . The method of  claim 1 , wherein the bridging oligonucleotide and the capture oligonucleotide are contained in a hairpin structure. 
     
     
         3 . The method of  claim 1 , wherein following ligation, uncleaved probe is removed from the surface.