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US 2021/0172014 A1Published application
Compositions and methods for improving sample identification in indexed nucleic acid libraries
AssigneesIllumina, Inc., Illumina Cambridge Limited
InventorsEric Hans Vermaas, Mahdieh Khosroheidari, Claire Bevis-Mott
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Abstract
The present invention is concerned with compositions and methods for improving the rate of correct sample identification in indexed nucleic acid library preparations for multiplex next generation sequencing by modifying or blocking 5′ and 3′ ends of pooled indexed polynucleotides from multiple samples, with an optional exonuclease treatment, prior to amplification and sequencing.
Claims21
1-21. (canceled)
A polynucleotide prepared for sequencing, comprising
- an adapter-target-adapter sequence,
- wherein the adapter sequence comprises a library specific sequence,
- wherein the polynucleotide is double stranded in a region comprising the target and at least a portion of the adapter on both ends of the target, and
- wherein 5′ and 3′ ends of the polynucleotide in a region of the adapter sequence are single stranded,
- wherein the 5′ ends are modified to prevent digestion by an enzyme having 5′ exonuclease activity, and
- wherein the 3′ ends are modified to inhibit digestion by an enzyme having 3′ exonuclease activity, to inhibit addition of nucleotides to the 3′ end by an enzyme having polymerase activity, or to inhibit digestion by an enzyme having 3′ exonuclease activity and to inhibit addition of nucleotides to the 3′ end by an enzyme having polymerase activity.
3 more elements
Dependencies are inferred from the claim text.