Method for highly sensitive nucleic acid detection using nanopore and non-specific nucleic acid binding agent

Abstract

Provided is a method of sensitively detecting nucleic acids in a nucleic acid sample, the method comprising: contacting the sample comprising the nucleic acid with a non-specific nucleic acid binding agent in an electrically conductive fluid medium; contacting the sample comprising the nucleic acid bound to the agent with a nanopore; and applying a voltage to the nanopore and monitoring a current change through the nanopore. The nucleic acid can be sensitively detected because a change in current amplitude through the nanopore is greater than when nucleic aid detection is performed without using an intercalator.

1 . An apparatus for detecting nucleic acids, the apparatus comprising:
 a nanopore;   a non-specific nucleic acid binding agent;   a device applying a voltage across the nanopore; and   a detector monitoring a current change through the nanopore.   
     
     
         2 . The apparatus of  claim 1 , wherein the non-specific nucleic acid binding agent is selected from the group consisting of an intercalator, a major groove binding agent and a minor groove binding agent. 
     
     
         3 . The apparatus of  claim 2 , wherein the intercalator is selected from the group consisting of YOYO (1,1′-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[(3-methyl-2(3H)-benzoxazolylide ne)methyl]]-,tetraiodide), TOTO (1-1′-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[(3-methyl-2(3H)-benzot hiazolylidene)methyl]]-,tetraiodide) and Ethidium Bromide (EtBr). 
     
     
         4 . The apparatus of  claim 1 , wherein the nanopore has a diameter ranging from 1.5 nm to 120 nm. 
     
     
         5 . The apparatus of  claim 1 , wherein the voltage ranges from 100 mV to 1000 mV.