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US 12,247,254 B2Granted
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.
Claims17
A polynucleotide prepared for sequencing, comprising
- an adapter-target-adapter sequence,
- wherein the adapter sequence comprises a library specific sequence,
- wherein the polynucleotide comprises a double stranded region,
- wherein the 5′ ends are each modified to prevent digestion by an enzyme having 5′ exonuclease activity,
- wherein the 3′ ends are each modified to inhibit digestion by an enzyme having 3′ exonuclease activity, to inhibit addition of nucleotides to the 3′ ends 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′ ends by an enzyme having polymerase activity, and
- wherein the 3′ ends each comprise a dideoxynucleotide.
3 more elements
Dependencies are inferred from the claim text.