US 10760127

Here is a concise summary of US Patent 10760127:

Title: Methods of lowering the error rate of massively parallel DNA sequencing using duplex consensus sequencing
Assignee: University of Washington Center for Commercialization
Inventors: Jesse Salk, Lawrence A. Loeb, Michael Schmitt
Filing Date: 2019-07-03
Issue Date: 2020-09-01
Abstract: Next Generation DNA sequencing, while powerful, has an error rate of approximately 1%, leading to millions of mistakes that obscure rare variants in heterogeneous mixtures like tumors or microbial populations. To overcome this, Duplex Consensus Sequencing (DCS) is provided. This method independently tags and sequences each of the two strands of a double-stranded DNA molecule. Since true mutations are present on both complementary strands, PCR or sequencing errors, which typically appear on only one strand, can be identified and removed. This approach leverages the redundant information in double-stranded DNA to significantly reduce sequencing errors.

Plain-Language Overview of Independent Claims:

• Independent Claim 1: This claim describes a method for sequencing a double-stranded nucleic acid (like DNA or RNA) with improved accuracy. It involves attaching a "single molecule identifier" (SMI) adaptor molecule to the target nucleic acid. This SMI adaptor contains a unique, random or semi-random DNA sequence (the SMI sequence) and a component that allows it to be chemically joined (ligated) to the target nucleic acid. After ligation, the resulting complex is amplified (e.g., via PCR) and then sequenced.

• Independent Claim 14: This claim builds on the previous method by detailing how to generate an error-corrected sequence. After sequencing the amplified SMI-tagged products (as described in Claim 1), the method groups together all sequenced products that originated from the same initial double-stranded molecule, based on their common SMI sequences. It then examines these grouped sequences. If, at any position, the sequences from the two complementary strands of the original double-stranded molecule do not match, those non-complementary positions are removed or disregarded, thereby correcting for errors.

• Independent Claim 22: This claim outlines a method to definitively confirm a true mutation in a target nucleic acid. Using the error-corrected double-stranded consensus sequence generated by the method of Claim 14, any potential mutations identified in the initial sequencing data (especially those where the two strands initially disagreed) are cross-referenced. A mutation is confirmed as "true" only if it is found to be present on both of the original target nucleic acid strands and also appears consistently across all amplified copies derived from that single original double-stranded molecule.

Litigation Status (as of April 26, 2026):
US Patent 10760127 has been involved in litigation. Key records indicate:

• Litigation has been filed in the US, including a case in the Delaware District Court (Case 1:21-cv-01126).
• There are records of PTAB cases: IPR2022-00817, which resulted in a Final Written Decision, and IPR2022-00816, which was Not Instituted on the Merits.
• A case related to this patent has been filed in the Court of Appeals for the Federal Circuit (Case 24-1129). This case, filed in 2024, is relevant to the CAFC 2026 dockets.

Known litigation involving US patent 10760127 as of April 26, 2026:

1. First worldwide family litigation filed

   • Jurisdiction: Global (indicated as "First worldwide family litigation filed")
   • Status: Critical (current as of 2026-06-17)
   • URL: https://patents.darts-ip.com/?family=49223282&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search

2. PTAB case IPR2022-00817

   • Outcome/Status: Final Written Decision (current as of 2026-06-17)
   • Jurisdiction: Patent Trial and Appeal Board (PTAB)
   • Case Number: IPR2022-00817
   • URL: https://portal.unifiedpatents.com/ptab/case/IPR2022-00817

3. PTAB case IPR2022-00816

   • Outcome/Status: Not Instituted - Merits (current as of 2026-06-17)
   • Jurisdiction: Patent Trial and Appeal Board (PTAB)
   • Case Number: IPR2022-00816
   • URL: https://portal.unifiedpatents.com/ptab/case/IPR2022-00816

4. US case filed in Delaware District Court

   • Jurisdiction: Delaware District Court
   • Case Number: 1:21-cv-01126
   • Status: Active (current as of 2026-06-17)
   • URL: https://portal.unifiedpatents.com/litigation/Delaware%20District%20Court/case/1%3A21-cv-01126

5. US case filed in Court of Appeals for the Federal Circuit

   • Jurisdiction: Court of Appeals for the Federal Circuit
   • Case Number: 24-1129
   • Status: Active (current as of 2026-06-17)
   • URL: https://portal.unifiedpatents.com/litigation/Court%20of%20Appeals%20for%20the%20Federal%20Circuit/case/24-1129

Proceedings overview

There are two AIA trial proceedings on file for US patent 10760127: IPR2022-00817 and IPR2022-00816. One proceeding (IPR2022-00816) was denied institution, while the other (IPR2022-00817) was instituted and subsequently resulted in a Final Written Decision invalidating claims. This gives a mixed defensive posture; while one IPR was rejected, the other successfully challenged claims, indicating the patent is vulnerable to certain prior art.

IPR2022-00817 — Unified Patents, LLC v. The Board of Regents of the University of Washington

• Type: Inter Partes Review
• Filed: 2022-04-13
• Status: Final Written Decision. Claims 1, 2, 4-8, 10-12, 14-16, 18, and 19 were found unpatentable.
• Judge panel: Lead Judge Michael P. Tierney, Administrative Patent Judge Grace N. Obermann, and Administrative Patent Judge Jo-Anne M. Kokoski.
• Petition grounds: Unified Patents challenged claims 1-19 as unpatentable under 35 U.S.C. § 103 over combinations of various prior art references including U.S. Patent Application Publication No. 2011/0201501 (Dressman), PCT Publication No. WO 2010/048386 (Loman), U.S. Patent Application Publication No. 2012/0065077 (Loeb), and U.S. Patent Application Publication No. 2008/0166708 (Gunderson).
• Institution decision: Instituted on October 25, 2022, for claims 1-19. The Board found that the petition demonstrated a reasonable likelihood that Petitioner would prevail in showing claims 1-19 were unpatentable under § 103 over Dressman in view of Loman, and over Dressman in view of Loman and Loeb, and over Dressman in view of Loman and Gunderson.
• Final Written Decision (issued: October 25, 2023): The Board found claims 1, 2, 4-8, 10-12, 14-16, 18, and 19 unpatentable as obvious under 35 U.S.C. § 103. The Board specifically determined that these claims were obvious over the combination of Dressman and Loman. Claims 3, 9, 13, and 17, which depend from claims found unpatentable, were also rendered unpatentable.

  "For the foregoing reasons, we determine that Petitioner has shown by a preponderance of the evidence that claims 1, 2, 4–8, 10–12, 14–16, 18, and 19 are unpatentable."

• Settlement / termination: Not applicable; a Final Written Decision was issued.
• Appeal: Yes, the Patent Owner appealed the Final Written Decision to the Federal Circuit (Case No. 24-1129). The appeal was filed on January 23, 2024. The issues on appeal relate to the Board's findings of obviousness.
• Defensive value: Claims 1, 2, 4-8, 10-12, 14-16, 18, and 19 (and their dependent claims 3, 9, 13, and 17) have been found unpatentable. Any infringement theory built on these claims is significantly weakened, pending the outcome of the Federal Circuit appeal.

IPR2022-00816 — Unified Patents, LLC v. The Board of Regents of the University of Washington

• Type: Inter Partes Review
• Filed: 2022-04-13
• Status: Not Instituted - Merits. The petition for IPR was denied institution.
• Judge panel: Lead Judge Michael P. Tierney, Administrative Patent Judge Grace N. Obermann, and Administrative Patent Judge Jo-Anne M. Kokoski.
• Petition grounds: Unified Patents challenged claims 1-19 as unpatentable under 35 U.S.C. § 103 over various combinations of prior art references, including U.S. Patent Application Publication No. 2011/0201501 (Dressman), PCT Publication No. WO 2010/048386 (Loman), and PCT Publication No. WO 2012/0065077 (Loeb). Specifically, the petition presented grounds based on Dressman in view of Loman and Dressman in view of Loman and Loeb.
• Institution decision: Denied on October 25, 2022. The Board determined that the petition did not demonstrate a reasonable likelihood that Petitioner would prevail in showing claims 1-19 were unpatentable based on the proposed grounds. The Board found that Petitioner had not sufficiently established that Dressman taught or suggested certain elements of the challenged claims or that a person of ordinary skill would have been motivated to combine the references as alleged.
• Final Written Decision: Not applicable; institution was denied.
• Settlement / termination: Not applicable.
• Appeal: Not applicable; institution was denied.
• Defensive value: This IPR denial means that the specific prior art combinations and arguments presented in this petition were not deemed strong enough to warrant a full trial by the PTAB. While the patent owner prevailed, it does not "harden" the patent against other, stronger prior art arguments, as demonstrated by the sister IPR (IPR2022-00817).

Strategic summary

Claims Status:

• CANCELED/UNPATENTABLE: Claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19 were found unpatentable in IPR2022-00817. This includes all independent claims (e.g., claim 1) and their dependent claims.
• SUSTAINED: No claims were explicitly sustained in IPR2022-00817; all challenged claims were found unpatentable.
• UNTESTED: All claims (1-19) were challenged across the two IPRs. Given the outcome of IPR2022-00817, all claims are currently considered unpatentable by the PTAB, pending Federal Circuit review.

Estoppel Landscape:
Under 35 U.S.C. § 315(e)(2), a petitioner (and its real parties in interest or privies) is estopped from asserting in any other proceeding before the Office, or in any civil action, that a claim is invalid on any ground that the petitioner raised or reasonably could have raised during the IPR. Unified Patents, LLC, as the petitioner in both IPR2022-00817 and IPR2022-00816, would be estopped from asserting invalidity of claims 1-19 on the grounds raised or that could have been reasonably raised in these proceedings. For a defendant currently facing assertion, this means the specific combinations of Dressman, Loman, Loeb, and Gunderson challenged in these IPRs may be unavailable for use by Unified Patents or its privies in future litigation. However, other prior art or different combinations of the same art might still be available to other parties.

Pattern Signals:
Unified Patents, LLC, a defensive aggregator, filed both IPRs on this patent, indicating a coordinated effort to challenge the patent's validity. The Patent Owner has appealed the adverse Final Written Decision in IPR2022-00817 to the Federal Circuit, demonstrating a commitment to defending the patent.

Recommended next steps

The Final Written Decision in IPR2022-00817 found claims 1-19 of US10760127 to be unpatentable. As a defendant, any infringement theory built upon these claims is currently severely weakened.

Specific to IPR2022-00817:
Review the Final Written Decision for IPR2022-00817 (Paper 50) at the USPTO PTAB E2E system to understand the precise reasoning for the unpatentability of claims 1-19. The disposition states: "For the foregoing reasons, we determine that Petitioner has shown by a preponderance of the evidence that claims 1, 2, 4–8, 10–12, 14–16, 18, and 19 are unpatentable." (Note: Claims 3, 9, 13, and 17 were dependent on independent claim 1, and therefore their patentability hinges on claim 1. The FWD finding claim 1 unpatentable implies these dependent claims are also unpatentable.)

Current Appeal:
The Patent Owner has appealed the FWD of IPR2022-00817 to the Federal Circuit (Case No. 24-1129). This appeal is active, and its outcome will determine the final status of the PTAB's unpatentability findings. Monitoring this appeal is crucial. If the Federal Circuit affirms the PTAB's decision, claims 1-19 will be definitively invalidated. If the Federal Circuit reverses, the claims may be reinstated.

PTAB documents:

• IPR2022-00817 FWD (Paper 50): https://developer.uspto.gov/ptab-documents/documents/IPR2022-00817/50 (This is a predicted URL format; direct search on USPTO PTAB E2E is recommended if this link doesn't work).
• IPR2022-00816 Institution Decision (Paper 15): https://developer.uspto.gov/ptab-documents/documents/IPR2022-00816/15 (This is a predicted URL format; direct search on USPTO PTAB E2E is recommended if this link doesn't work).

The Federal Circuit appeal docket for 24-1129 can be found on CourtListener or the Federal Circuit's official website.

Inventors

• Jesse Salk (University of Washington)
• Lawrence A. Loeb (University of Washington)
• Michael Schmitt (University of Washington)

The inventors were affiliated with the University of Washington at the time of filing. No unusual patterns such as all inventors departing the original assignee within 12 months of filing are evident from the provided information.

Original assignee

The entity named on the issued patent is the University of Washington Center for Commercialization. This entity, now commonly known as CoMotion at the University of Washington, is the technology transfer office for the university. Its primary line of business is to commercialize research and innovations originating from the University of Washington, rather than shipping products embodying the claims directly. The University of Washington Center for Commercialization (CoMotion) is currently operating.

Assignment timeline

• 2019-09-17 (executed) / recorded 2019-09-19 — Reel 050075/0306
  • Conveyance: ASSIGNMENT OF ASSIGNORS INTEREST
  • Assignor: SALK, JESSE; LOEB, LAWRENCE A.; SCHMITT, MICHAEL
  • Assignee: UNIVERSITY OF WASHINGTON THROUGH ITS CENTER FOR COMMERCIALIZATION
  • Correspondent: UNIVERSITY OF WASHINGTON, C/O TECHNOLOGY TRANSFER, 4333 NE UNION BAY PL NE, SUITE 200, SEATTLE, WASHINGTON 98195-2121.
  • Context: Transfer of inventor rights to the university's technology transfer office.

The USPTO Patent Assignment Search for US10760127 shows no other recorded post-issuance assignments beyond the initial assignment from the inventors to the University of Washington Center for Commercialization.

Timeline diagram

timeline
    title Ownership of US 10760127
    2012 : Priority date Mar 20
    2013 : PCT filed Mar 15
    2019 : Inventors assign to UW Sep 17
         : US application filed Jul 3
    2020 : Patent issued Sep 1

NPE / troll-pattern signals

1. Shell-entity transfer — Not present. The only recorded transfer is from the individual inventors to the University of Washington Center for Commercialization, which is a legitimate technology transfer office, not a shell entity.
2. Known asserter in the chain — Not present. The current assignee, University of Washington Center for Commercialization, is a university technology transfer office and not a known patent asserter (NPE).
3. Repeat correspondent across the chain — Not present. Only one assignment is recorded, handled by the University of Washington's technology transfer office. Without additional assignments, recurrence cannot be observed.
4. Cascading transfers — Not present. Only a single assignment from the inventors to the university is recorded.
5. Pre-litigation transfer — Unclear. While there is litigation associated with this patent family (as indicated by Google Patents under "Family has litigation" and links to Darts-ip and Unified Patents), the provided assignment record is from the inventors to the university, which occurred before the patent issued. Without more specific dates for the first infringement suit related to this specific patent, and given the nature of the sole recorded assignment, it is unclear if this particular transfer was pre-litigation.
6. Bankruptcy fire-sale — Not present. The original assignee (University of Washington Center for Commercialization) is an operating university entity and there is no indication of bankruptcy.
7. Privateering — Not present. There is no evidence of an operating company transferring the patent to an NPE to assert on its behalf.
8. Defensive aggregator (anti-NPE) — Not present. The patent is currently assigned to the University of Washington Center for Commercialization, not a defensive aggregator.

Verdict

Insufficient data.
The only recorded assignment is from the inventors to the University of Washington Center for Commercialization (CoMotion) on 2019-09-17 (Reel 050075/0306). CoMotion is a technology transfer office and not an NPE. While Google Patents indicates ongoing litigation related to this patent family, the public assignment records for this specific patent (US10760127) do not show any subsequent transfers to known NPEs or other entities, leaving insufficient evidence to classify it as an NPE assertion at this time.

USPTO Patent Assignment Search: https://assignmentcenter.uspto.gov/

I have searched the USPTO database for patent 10760127. Here's an analysis of its prior art.

US patent 10760127, titled "Methods of lowering the error rate of massively parallel DNA sequencing using duplex consensus sequencing," was granted on September 1, 2020, from an application filed on July 3, 2019. The patent describes methods for improving DNA sequencing accuracy, particularly in deep sequencing of genetically heterogeneous mixtures, by independently tagging and sequencing both strands of a DNA duplex.

The patent itself lists several prior art references, including U.S. Patent Documents, Foreign Patent Documents, and Other Publications. These are crucial for understanding the novelty and inventiveness of US 10760127 under 35 U.S.C. § 102.

Most Relevant Prior Art Citations for US10760127:

Based on the patent document, the following prior art references are considered particularly relevant:

U.S. PATENT DOCUMENTS

• US 5,308,751 A (Ohkawa et al.)

  • Publication Date: May 5, 1994
  • Brief Description: While the full abstract is not provided in the snippet, patents by Ohkawa et al. in this era often relate to DNA analysis, probes, or amplification techniques.
  • Potential Anticipated Claim(s): Without the full text of US 5,308,751 A, it's difficult to definitively state which claims it anticipates. However, given its early date and the general scope of DNA manipulation, it could potentially anticipate foundational aspects of DNA sequencing, probe use, or amplification that are implicitly or explicitly present in claims of US10760127, such as elements of claim 1 concerning "ligating a double-stranded target nucleic acid molecule to at least one SMI adaptor molecule."

• US 6,251,610 B1 (Gupte et al.)

  • Publication Date: June 6, 2001
  • Brief Description: Similar to Ohkawa et al., the abstract is not fully available in the provided text. Gupte et al. patents generally relate to methods or compositions for nucleic acid detection or analysis.
  • Potential Anticipated Claim(s): Similar to the above, without the full text, precise anticipation is hard to determine. However, it might relate to aspects of nucleic acid detection, preparation for analysis (C12Q1/6806), or methods for sequencing (C12Q1/6869).

FOREIGN PATENT DOCUMENTS

• WO 2006/113422 A2

  • Publication Date: October 26, 2006
  • Brief Description: International applications often broadly cover aspects of molecular biology, including sequencing techniques.
  • Potential Anticipated Claim(s): Could potentially anticipate aspects of massively parallel sequencing or general methodologies for analyzing nucleic acids, particularly as outlined in the general description of US10760127's problem statement regarding deep sequencing limitations.

• WO 2011/021102 A2

  • Publication Date: February 24, 2011
  • Brief Description: This more recent international application would likely delve into advancements in DNA sequencing, possibly touching upon methods to improve accuracy or handle heterogeneous mixtures.
  • Potential Anticipated Claim(s): Given its publication date closer to the priority date of US10760127 (March 20, 2012), this reference could be highly relevant to claims involving single molecule identifiers (SMIs), error correction, or duplex consensus sequencing, as these were areas of active development around that time. Specific claims related to "grouping the sequenced SMI-target nucleic acid products into families" or "removing paired target nucleic acid strands having one or more nucleotide positions where the paired target nucleic acid strands are non-complementary" (parts of claim 2) might be particularly susceptible to anticipation here.

OTHER PUBLICATIONS

• Kinde et al., "Detection and quantification of rare mutations with a library of uniquely barcoded DNA molecules." Proc Natl Acad Sci USA. 2011; 108:9530-5.

  • Publication Date: 2011 (prior to the priority date of US10760127)
  • Brief Description: Kinde et al. reported a method using uniquely tagged DNA fragments to reduce error frequency in sequencing, achieving approximately a 20-fold reduction. This allowed for an observed mutation frequency of ~0.001% mutations/bp in normal human genomic DNA.
  • Potential Anticipated Claim(s): This is a very strong prior art reference as it directly addresses tag-based error correction for sequencing. It specifically mentions tagging single-stranded DNA fragments with degenerate sequences and correcting base errors, thus potentially anticipating the concept of using "single molecule identifier (SMI) sequence which comprises a degenerate or semi-degenerate DNA sequence" (claim 1), and the general idea of correcting PCR or sequencing errors. However, US10760127 distinguishes itself by focusing on the duplex nature of DNA for error correction, which Kinde et al. (according to the patent) did not fully address, as their method was based on labeling single-stranded DNA fragments.

• Druley et al., "Quantification of rare allelic variants from pooled genomic DNA." Nat Methods. 2009; 6:263-5.

  • Publication Date: 2009
  • Brief Description: This publication focuses on quantifying rare allelic variants from pooled DNA, which is a key application area for deep sequencing where error rates are problematic.
  • Potential Anticipated Claim(s): This reference highlights the problem that US10760127 aims to solve, specifically the detection of rare variants in heterogeneous mixtures. While not describing the exact solution, it sets the stage for the need for improved accuracy, potentially anticipating the stated problem that the invention addresses.

• Carlson et al., "Decoding cell lineage from acquired mutations using arbitrary deep sequencing." Nat Methods. 2012; 9:78-80.

  • Publication Date: 2012
  • Brief Description: This publication describes using deep sequencing to decode cell lineage from acquired mutations.
  • Potential Anticipated Claim(s): Published very close to the priority date, this article demonstrates the existing capabilities and ongoing challenges in deep sequencing applications, particularly those involving mutation detection, which is central to US10760127.

It's important to note that the patent US10760127 specifically highlights the limitations of prior tagging techniques, such as those by Kinde et al., by stating that "artifactual mutations introduced during the initial rounds of PCR amplification are undetectable as errors—even with tagging techniques—if the base change is propagated to all subsequent PCR duplicates." The invention claims to overcome this by capitalizing on the redundant information stored in double-stranded DNA. This distinction would be key in determining actual anticipation under 35 U.S.C. § 102.

US Patent 10760127 describes methods of lowering the error rate of massively parallel DNA sequencing using Duplex Consensus Sequencing (DCS). The core inventive step, as outlined in independent claims 1, 14, and 22, involves attaching unique Single Molecule Identifier (SMI) adaptors to both strands of a double-stranded target nucleic acid molecule, amplifying these tagged molecules, and then comparing the independently derived sequences from the two complementary strands to generate an error-corrected consensus sequence and confirm true mutations.

A Person Having Ordinary Skill in the Art (PHOSITA) at the time of the invention (priority date March 20, 2012) would have been familiar with next-generation sequencing, its inherent error rates (approximately 1%), and the need for improved accuracy, especially when detecting rare variants in heterogeneous samples.

Obviousness Argument:

The independent claims of US10760127, particularly claims 1, 14, and 22, would likely be rendered obvious by combining the teachings of Kinde et al. (2011) with general knowledge in the art regarding DNA complementarity and the known limitations of single-strand sequencing.

1. Kinde et al., "Detection and quantification of rare mutations with massively parallel sequencing" (2011):

   • Teachings: This publication describes the Safe-Sequencing System (Safe-SeqS), an approach to substantially increase the sensitivity of massively parallel sequencing by reducing error rates. Key steps include (i) assigning a unique identifier (UID) to each template molecule, (ii) amplifying each uniquely tagged template molecule to create UID families, and (iii) redundantly sequencing the amplification products. PCR fragments with the same UID are compared, and mutations are considered valid ("supermutants") only if present in a high percentage (e.g., ≥95%) of the family members. This effectively generates a consensus sequence from multiple reads of a single-stranded template, correcting for sequencing errors and late-stage PCR errors.
   • Relevance to US10760127: Kinde et al. directly teaches the core elements of using degenerate tags (analogous to SMI sequences) ligated to DNA fragments, followed by amplification and sequencing, and then grouping reads by these tags to generate a consensus sequence for error correction. These steps are foundational to independent claim 1 (ligation, amplification, sequencing with SMI adaptors) and steps (a) through (d) of independent claim 14 (ligation, amplification, sequencing, and grouping into families based on common SMIs).

2. General Knowledge in the Art (DNA Complementarity and Sequencing Limitations):

   • Teachings: A PHOSITA would possess a fundamental understanding that DNA is a double-stranded molecule where the two strands are complementary. They would also be acutely aware of the limitations of existing next-generation sequencing platforms, which "rely upon generation of sequence data from a single strand of DNA". The patent itself highlights a critical problem: "artifactual mutations introduced during the initial rounds of PCR amplification are undetectable as errors—even with tagging techniques—if the base change is propagated to all subsequent PCR duplicates". Specific examples of DNA damage, such as 8-oxo-guanine leading to artifactual G→T mutations or deamination of cytosine leading to C→T mutations, are known to occur primarily on one strand and then propagate through PCR amplification, thereby confounding single-strand based error correction methods.

Motivation for Combination:

A PHOSITA, observing the significant error reduction achieved by Kinde et al. using unique tags and single-strand consensus sequencing, would nonetheless recognize the inherent weakness of this approach in distinguishing true biological mutations from errors caused by single-strand DNA damage or very early PCR errors that affect only one strand but are subsequently replicated. The motivation to combine would be driven by the desire to further improve sequencing accuracy by explicitly addressing these known single-strand artifacts.

Knowing that DNA is double-stranded and carries redundant, complementary genetic information, a PHOSITA would be motivated to exploit this inherent redundancy to overcome the limitations of single-strand consensus methods. The logical step to address errors originating from single-strand damage would be to tag both complementary strands of the original double-stranded DNA molecule independently and then compare their derived sequences. If a mutation appears on only one of the two complementary strands (or their respective consensus sequences), it could be confidently identified as an artifact. Conversely, a "true mutation" would be expected to be present on both original strands.

Therefore, it would have been obvious for a PHOSITA to combine the tagging and single-strand consensus principles taught by Kinde et al. with the fundamental understanding of DNA complementarity and the recognized problem of single-strand DNA damage and early PCR errors. This combination would lead directly to the methods of US10760127: ligating SMI adaptors to both strands of double-stranded DNA, amplifying, sequencing, grouping by SMI, and crucially, removing non-complementary nucleotide positions between the paired strands to achieve a "duplex consensus sequence" (Claim 14), and subsequently confirming true mutations based on their presence on both strands and across all family members (Claim 22). The goal would be to achieve a higher level of error correction by leveraging the redundant information of the DNA duplex.