Systems for nucleic acid sequencing

Abstract

Systems for nucleic acid sequencing include template constructs that comprise double stranded portions in a partially or completely contiguous constructs, to provide for redundant sequence determination through sequencing of both sense and antisense strands.

What is claimed is: 
     
         1 . A system for sequencing template nucleic acid molecules with improved accuracy, comprising:
 a plurality of template nucleic acid molecules comprising a double-stranded portion, the double-stranded portion consisting of a forward strand and a reverse strand, wherein the forward strand and the reverse strand are fully complementary to each other, and a linking oligonucleotide that links a 3′ end of the forward strand to a 5′ end of the reverse strand of each of said template nucleic acid molecules;   a sequencing instrument in contact with the plurality of template nucleic acid molecules that performs at least one single-molecule sequencing process on each of said plurality of template molecules, wherein the forward strand and the reverse strand in each of said plurality of template molecules are fully complementary to each other, each of said at least one single-molecule sequencing process consisting of sequencing only one single template molecule of said plurality of template molecules, thereby generating a sequence read from each of said plurality of template molecules, said sequence read comprising sequences of both the forward strand and the reverse strand of each of said plurality of template molecules; and   a computer that determines the sequence of each of said plurality of template molecules by analyzing said sequence read from each of said plurality of template molecules, including comparing the sequences of the forward strand and the reverse strand from each of said plurality of template molecules, thereby improving sequencing accuracy of said nucleic acid molecules.   
     
     
         2 . The system of  claim 1 , wherein each of said plurality of template molecules comprises a primer sequence that is complementary to a single-stranded portion of said plurality of template molecules. 
     
     
         3 . The system of  claim 1 , wherein said at least one single-molecule sequencing process comprises initiating sequencing at a gap within each of said plurality of template molecules. 
     
     
         4 . The system of  claim 1 , wherein said at least one single-molecule sequencing process includes single-molecule sequencing of the linking oligonucleotide. 
     
     
         5 . The system of  claim 1 , wherein said at least one single-molecule sequencing process is performed using an electrochemical system. 
     
     
         6 . The system of  claim 1 , wherein said at least one single-molecule sequencing process is performed using a nanopore sensor. 
     
     
         7 . The system of  claim 1 , wherein said at least one single-molecule sequencing process is performed using a sequencing by synthesis technology. 
     
     
         8 . The system of  claim 1 , wherein said double-stranded portion comprises at least 500 base pairs. 
     
     
         9 . The system of  claim 1 , wherein said linking oligonucleotide comprises a registration sequence.