Methods for indexing samples and sequencing multiple polynucleotide templates
AssigneeIllumina Cambridge Limited
InventorsHelen Bignell, Louise Fraser, Niall Anthony Gormley
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Abstract
The invention relates to methods for indexing samples during the sequencing of polynucleotide templates, resulting in the attachment of tags specific to the source of each nucleic acid sample such that after a sequencing run, both the source and sequence of each polynucleotide can be determined. Thus, the present invention pertains to analysis of complex genomes (e.g., human genomes), as well as multiplexing less complex genomes, such as those of bacteria, viruses, mitochondria, and the like.
Claims17
A method for preparing a nucleic acid library from a plurality of subsets of double stranded nucleic acids, wherein each subset is obtained from a different source, comprising:
- (a) providing the plurality of subsets of double stranded nucleic acids;
- (b) ligating a tagged adaptor to each end of the double stranded nucleic acids to generate adaptor-target-adaptors, wherein the tagged adaptor comprises at least one region comprising a tag sequence that differentiates adaptor-target-adaptors from different subsets;
- (c) amplifying the adaptor-target-adaptors of each subset with a pair of amplification primers complementary to the tag sequence to generate amplified adaptor-target-adaptors from each subset;
- (d) pooling the amplified adaptor-target-adaptors of the plurality of subsets;
- and
- (e) immobilizing the pooled adaptor-target-adaptors on a surface.
Dependencies are inferred from the claim text.