Paired-end reads in sequencing by synthesis
AssigneeHelicos BioSciences Corporation
InventorTimothy D. Harris
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Abstract
The disclosure provides methods of generating paired reads in sequencing-by-synthesis process, particularly, in systems with relatively short read lengths (e.g., 15-35 bases), such as for example, in single molecule sequencing by synthesis. Several implementations of the methods are provided. Of particular advantage are the methods that permit re-sequencing of the template, which yields lower error rates. The invention further provides methods of using paired reads, for example, for positioning them over repeats or for assembly into large sequences, including whole genome assembly.
Claims20
1-26. (canceled)
A method for generating paired reads from a strand of a nucleic acid duplex, the method comprising:
- a) providing a nucleic acid template;
- b) conducting a sequencing-by-synthesis reaction using detectably labeled nucleotides to obtain a first read of the template;
- and
- c) conducting a sequencing-by-synthesis reaction using detectably labeled nucleotides to obtain a second read of the template, wherein the second read is separated from the first read along the length of the template.
A method for generating paired reads from a strand of a nucleic acid duplex, the method comprising:
- a) providing a nucleic acid template;
- b) conducting a sequencing-by-synthesis reaction using detectably labeled nucleotides to obtain a first read of the template;
- c) synthesizing a spacer, wherein the synthesized spacer comprises unlabeled nucleotides;
- and
- d) conducting a sequencing-by-synthesis reaction using detectably labeled nucleotides to obtain a second read of the template, wherein the second read is separated from the first read by the spacer.
A method for generating paired reads from a strand of a nucleic acid duplex, the method comprising:
- a) providing a nucleic acid template, wherein the template is indirectly associated with a solid support;
- b) conducting a sequencing-by-synthesis reaction using detectably labeled nucleotides to obtain a first read of the template;
- and
- c) conducting a sequencing-by-synthesis reaction using detectably labeled nucleotides to obtain a second read of the template, wherein the second read is separated from the first read along the length of the template.
Dependencies are inferred from the claim text.