Sensing apparatus for amplification and sequencing of template polynucleotides and array for amplification of template polynucleotides

Abstract

Provided is a sensing apparatus comprising a chip for integrated amplification and sequencing of a template polynucleotide in a sample. The apparatus comprises a chip with at least one ISFET in a well or chamber, amplification means for amplifying the template polynucleotide on a surface of said chip and comprising at least one heating means suitable for conducting amplification of the template polynucleotide at temperatures elevated with respect to room temperature, and sequencing means for sequencing the amplified template polynucleotide in said well or chamber. Methods of use are also provided.

1 . An array for amplification and sequencing of template polynucleotides, the array comprising:
 a plurality of wells wherein each well is exposed to an ISFET;   a plurality of capture beads, wherein each capture bead comprises a single template binding site,   wherein the wells and capture beads are relatively sized so that only one bead will fit into each well;   a removable seal arranged to cover the wells and isolate each well from adjacent wells; and   a heating element.   
     
     
         2 . The array according to  claim 1 , wherein each bead comprises a single immobilised oligonucleotide. 
     
     
         3 . The array according to  claim 2 , wherein the oligonucleotide is a capture site for capturing a template for sequencing. 
     
     
         4 . The array according to  claim 2 , wherein the beads are Streptavidin beads and the oligonucleotides are biotin-conjugated oligonucleotides. 
     
     
         5 . The array according to  claim 1 , wherein the beads are fabricated from any one of a group consisting of inorganics, natural polymers, and synthetic polymers. 
     
     
         6 . The array according to  claim 5 , wherein the beads are fabricated from any one of a group consisting of cellulose, cellulose derivatives, glass silica, cross-linked dextrans, agarose gel and polystyrene. 
     
     
         7 . The array according to  claim 6 , wherein the cross-linked dextrans is Sephadex (R). 
     
     
         8 . The array according to  claim 1 , wherein the beads are suitable for covalent attachment. 
     
     
         9 . The array according to  claim 1 , wherein the beads are magnetic. 
     
     
         10 . The array according to  claim 1 , wherein the beads are provided with chemical groups that are bound to the surface of the bead to enable the attachment of oligonucleotides. 
     
     
         11 . The array according to  claims 1 , wherein the removable seal is a liquid. 
     
     
         12 . The array according to claims,  1  wherein the removable seal is a flexible membrane. 
     
     
         13 . The array according to  claim 12 , wherein the seal comprises an adhesive. 
     
     
         14 . The array according to  claim 1 , wherein the heating element is a resistive heating element. 
     
     
         15 . The array according to  claim 1 , further comprising a temperature sensor. 
     
     
         16 . The array according to  claim 11 , wherein the removable seal is a mineral oil.