Methods for indexing samples and sequencing multiple polynucleotide templates
AssigneeIllumina Cambridge Limited
InventorsHelen Bignell, Louise Fraser, Niall Anthony Gormley
Life of this patent
Estimating the term…
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 system comprising adaptors for use in distinguishing a source of a nucleic acid in a nucleic acid library, wherein first and second adaptors each comprise a first strand and a second strand, wherein:
- a portion of the 3′ end of the first strand and a portion of the 5′ end of the second strand is double stranded and comprises a double stranded tag sequence, wherein the tag sequence of the first adaptor is different from the tag sequence of the second adaptor;
- a portion of the 5′ end of the first strand and a portion of the 3′ end of the second strand are each single stranded;
- the 5′ end of the first strand comprises a sequencing primer binding site;
- and
- the 3′ end of the second strand comprises a universal primer binding sequence or complement thereof.
A system to distinguish a source of a target nucleic acid in a nucleic acid library, comprising:
- (a) a first solution comprising:
- a first plurality of double stranded target nucleic acids, and
- a first plurality of forked adaptors;
- and
- (b) a second solution comprising:
- a second plurality of double stranded target nucleic acids, and
- a second plurality of forked adaptors;
- wherein each forked adaptor of the first and second plurality of forked adaptors comprises:
- a double-stranded tag sequence, wherein the tag sequence of the first plurality of forked adaptors is different from the tag sequence of the second plurality of forked adaptors;
- and
- a first strand and a second strand, wherein each strand has a 3′ end and a 5′ end, wherein a portion of the 3′ end of the first strand and a portion of the 5′ end of the second strand is double stranded and comprises the tag sequence, and a portion of the 5′ end of the first strand and a portion of the 3′ end of the second strand are each single stranded, wherein the 5′ end of the first strand comprises a sequencing primer binding site, and the 3′ end of the second strand comprises a universal primer binding sequence or complement thereof.
Dependencies are inferred from the claim text.