Reverse complement adapters for the mitigation of UMI hopping

Patent metadata

Field	Value
Publication number	US-11053537-B2
Application number	US-201815989784-A
Country	US
Kind code	B2
Filing date	May 25, 2018
Priority date	May 25, 2017
Publication date	Jul 6, 2021
Grant date	Jul 6, 2021

Abstract

The invention pertains to construction of next-generation DNA sequencing (NGS) libraries for whole genome sequencing, targeted resequencing, sequencing-based screening assays, metagenomics, or any other application requiring sample preparation for NGS.

First claim

What is claimed is:

1. A method of preparing a target nucleic acid fragment for sequencing, the method comprising:
a. ligating a partially duplexed adapter to the ends of the target nucleic acid wherein the partially duplexed adapter has a top strand and a bottom strand;
i. wherein the top strand comprises from the 5′ to 3′ direction a P7rc sequence, optionally a UMI sequence, optionally a sample index sequence and a sequence complementary to the bottom strand; and
ii. wherein the bottom strand comprises from the 3′ to 5′ direction a P5rc sequence, optionally a second UMI sequence, optionally a sample index sequence, and a sequence complementary to the top strand.

2. The method of claim 1 wherein the top strand sample index sequence and the bottom strand sample index sequence are different.

3. The method of claim 1 wherein the top strand sample index sequence and the bottom strand sample index sequence are the same.

4. A method of preparing a target nucleic acid fragment for sequencing, the method comprising:
a. ligating a partially duplexed adapter to the ends of the target nucleic acid wherein the partially duplexed adapter has a top strand and a bottom strand;
i. wherein the top strand comprises SEQ ID NO: 5 and wherein the bottom strand comprises SEQ ID NO: 6.

Assignees

• Integrated Dna Tech Inc

Inventors

• Jarosz Mirna
• Light Madelyn
• Wang Jiashi
• Giorda Kristina

Classifications

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q1/6855 Primary

  Ligating adaptors · CPC title

• C12N15/1065

  Preparation or screening of tagged libraries, e.g. tagged microorganisms by STM-mutagenesis, tagged polynucleotides, gene tags · CPC title

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• C12Q1/6848

  characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction · CPC title