Electron induced fluorescence for nucleic acid sequencing

Abstract

An apparatus, compositions and related methods for sequencing a target nucleic acid are described. In certain embodiments, the apparatus is a microfluidic apparatus comprising an input chamber, microchannel, output chamber and a detection unit that is operatively connected to the microchannel. In preferred embodiments, the methods include hybridizing a target nucleic acid to one or more probe libraries, moving the hybridized target nucleic acid past the detector, and detecting bound probes. Probe libraries may comprise oligonucleotides or oligonucleotide analogs, preferably with each probe uniquely labeled. A linear order of labeled probes hybridized to the target nucleic acid can be detected and the target nucleic acid sequence deduced. In preferred embodiments, probe labels are detected by analysis of electron-induced fluorescence of probes labeled with conductive polymers.

1 - 17  (Cancelled)  
     
     
         18 . A nucleic acid sequencing apparatus comprising: 
 a) an input chamber;    b) a microchannel in fluid communication with the input chamber; and    c) a detection unit operably coupled to the microchannel, the detection unit comprising an electron beam source and a detector.    
     
     
         19 . The apparatus of  claim 18 , further comprising an information processing and control system operably coupled to the detector.  
     
     
         20 . The apparatus of  claim 18 , further comprising a fluidic focusing region between the input chamber and the microchannel.  
     
     
         21 . The apparatus of  claim 18 , further comprising a target nucleic acid hybridized to multiple labeled probes, each probe distinguishably labeled.  
     
     
         22 . The apparatus of  claim 21 , wherein the probes are labeled with conductive polymer molecules.  
     
     
         23 . The apparatus of  claim 20 , wherein the fluidic focusing region contains a separating comb.  
     
     
         24 . The apparatus of  claim 18 , wherein the electron beam source is a micro column electron beam source.  
     
     
         25  (Cancelled)  
     
     
         26 . The apparatus of  claim 18 , further comprising an array of microfluidic sequencing units.