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US 11,697,836 B2Granted
Normalization of NGS library concentration
AssigneeIntegrated Dna Technologies, Inc.
InventorsVladimir Makarov, Sergey Chupreta
Life of this patent
Estimating the term…
Abstract
A bottleneck in the Next Generation Sequencing (NGS) workflow is the quantification of libraries for accurate pooling and loading of the sequencing instrument flow cell or chip. Disclosed herein are methods that improve performance and reduce time compared to existing methods.
Claims9
A kit comprising:
- a first primer comprising a first portion, a second portion and a third portion, wherein the first portion is located at a 3′ end of the first primer, wherein the second portion comprises 3 or more consecutive ribonucleotide bases and is located 5′ to the first portion, and wherein the third portion is located 5′ adjacent to the second portion and comprises two or more deoxynucleotides;
- a second primer;
- a ligase;
- a probe, wherein the probe comprises a modification to provide resistance to digestion by an enzyme with 3′-5′ exonuclease activity, and wherein at least a portion of the probe is complementary to at least a portion of the third portion of the first primer and to the three or more consecutive ribonucleotide bases;
- and
- an exonuclease.
3 more elements
A kit comprising:
- a first primer comprising a first portion, a second portion, and a third portion, wherein the first portion is located at a 3′ end of the first primer, wherein the second portion comprises 3 or more consecutive ribonucleotide bases and is located 5′ to the first portion, and wherein the third portion is located 5′ adjacent to the second portion and comprises a low complexity sequence selected from the group consisting of poly(A), poly(T), poly(G), poly(C), poly(AG), poly(AC), poly(GT), poly(CT), poly(AT), poly(GC), a trinucleotide, a tetranucleotide, and a pentanucleotide;
- a second primer;
- a ligase;
- a probe, wherein the probe comprises a modification to provide resistance to digestion by an enzyme with exonuclease activity;
- and
- an exonuclease.
3 more elements
Dependencies are inferred from the claim text.