Prosecution Insights
Last updated: April 19, 2026
Application No. 17/997,696

QUANTITATIVE BLOCKER DISPLACEMENT AMPLIFICATION (QBDA) SEQUENCING FOR CALIBRATION-FREE AND MULTIPLEXED VARIANT ALLELE FREQUENCY QUANTITATION

Non-Final OA §103§112§DP
Filed
Nov 01, 2022
Examiner
PRIEST, AARON A
Art Unit
1681
Tech Center
1600 — Biotechnology & Organic Chemistry
Assignee
William Marsh Rice University
OA Round
1 (Non-Final)
61%
Grant Probability
Moderate
1-2
OA Rounds
3y 0m
To Grant
87%
With Interview

Examiner Intelligence

Grants 61% of resolved cases
61%
Career Allow Rate
486 granted / 794 resolved
+1.2% vs TC avg
Strong +26% interview lift
Without
With
+26.0%
Interview Lift
resolved cases with interview
Typical timeline
3y 0m
Avg Prosecution
30 currently pending
Career history
824
Total Applications
across all art units

Statute-Specific Performance

§101
7.0%
-33.0% vs TC avg
§103
31.8%
-8.2% vs TC avg
§102
21.7%
-18.3% vs TC avg
§112
22.4%
-17.6% vs TC avg
Black line = Tech Center average estimate • Based on career data from 794 resolved cases

Office Action

§103 §112 §DP
Notice of Pre-AIA or AIA Status The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA . DETAILED ACTION Status of the Claims Claims 1-4, 6-9, 14-19, 21, 23-24, 27-28 and 34 are pending. Claims 1-4, 6, 8, 9 and 14-15 are the subject of this NON-FINAL Office Action. This is the first action on the merits. Election/Restrictions Applicant’s election without traverse of Group I (claims 1-4, 6-9, 14-15) and the oligo-primer configuration species of Figure 1 in the reply filed on 10/16/2025 is acknowledged. The elections read on claims 1-4, 6, 8, 9 and 14-15. Claims 5, 7, 16-19, 21, 23-24, 27-28 and 34 are withdrawn. Claim Rejections - 35 USC § 112- Indefiniteness The following is a quotation of 35 U.S.C. 112(b): (B) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention. Claim 1-4, 6, 8, 9 and 14-15 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor, or for pre-AIA the applicant regards as the invention. Claim 1 is confusing. First, as to the “fifth oligonucleotide,” it is unclear where this binds. As an initial matter, the parentheses are confusing because it is not clear if the phrase in the parentheses redefines, or adds limitations to the simple “fifth oligonucleotide” (“a fifth oligonucleotide (Blocker Displacement Amplification (BDA) forward primer)”). The same issue exists with the “sixth oligonucleotide” (“a sixth oligonucleotide (BDA blocker)”). Next, “a specific genomic region” that the “eighth region” of the “fifth oligonucleotide” “targets” is confusing. Claim 1 repeatedly recites numbered “specific genomic regions” (“first specific genomic region”; “second specific genomic region”), yet here Applicants use “a specific genomic region.” It is not clear if this “a specific genomic region” is one of the previously-recited “specific genomic regions” or a new one. The “fifth oligonucleotide” is described as including a “genomic region targeted by the eighth region,” yet again it is unclear which “genomic region” this means due to lack of antecedent basis. Finally, and in light of the above, it is unclear where this “fifth oligonucleotide” binds to the “genomic region.” The fifth oligo’s unclear “genomic region targeted by the eighth region is between 1 and 20 nucleotides closer to the seventh region compared to the genomic region targeted by the third region.” “[T]he genomic region targeted by the third region” lacks antecedent basis. Yet, beyond this problem, it is not clear how this closeness is measured if the “genomic region targeted by the eighth region” is on a different strand from the “seventh region” and/or “the genomic region targeted by the third region.” Even more, from what precise point is this closeness measured? For example, the 3’-end of each oligo? The 5’-end? A “region” is any size. How is one binding region (say, 20 bases) closer to another binding region (say 10 bases) than a second binding region (say 15 bases)? The skilled artisan is left to guess the metes and bounds. The ”sixth oligonucleotide” shares “4 or more nucleotides at the 3' end of the BDA forward primer sequence [which] are also present at or near the 5' end of the BDA blocker sequence,” which is confusing. “Near” is a subjective term of degree with no clear meaning. It is, in itself, vague. Are 10 bases away from the 5’-end “near” it? Twenty bases? There is simply no clear metes and bounds to this vague term. Claim Rejections - 35 USC § 103 The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action: A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made. Claim(s) 1-4, 6, 8, 9 and 14-15 is/are rejected under 35 U.S.C. 103 as being unpatentable over WOOD (US20210180121), in view of BIGLIA (US20160076089) as evidenced by US20170067090, US20140017685 and US20170233821. The prior art as a whole demonstrates that it would have been obvious to a skilled artisan at the time of filing to detect SNPs using known high-sensitivity assays such as competitive blocker amplifications to achieve greater detection sensitivity in sequencing assays with a reasonable expectation of success. As to claim 1, WOOD teaches a method for labeling and amplifying each strand of between 1 and 10,000 target genomic regions with an oligonucleotide barcode sequence by polymerase chain reaction (PCR), the method comprising: (a) introducing into a DNA sample comprising the between 1 and 10,000 target genomic regions, for each target genomic region (Fig. 5): (i) a first oligonucleotide F1/F2, comprising from 5′ to 3′ end, (A) a first region, (B) a second region with a length between 0 and 50 nucleotides, and (C) a third region targeting a first specific genomic region (Fig. 5, PCR I); and (ii) a second oligonucleotide R1/R2, comprising from 5′ to 3′ end, (A) a fourth region, (B) a fifth region with a length between 0 and 50 nucleotides, (C) a sixth region comprising a unique molecular identifier (UMI) comprising at least four degenerate nucleotides, and (D) a seventh region targeting a second specific genomic region (Fig. 5, PCR I); (b) performing at least two cycles of PCR amplification to generate a first PCR amplification product (2-3 cycles for first PCR in Fig. 5; paras. 0072-77); (c) introducing into the first PCR amplification product: (i) a third oligonucleotide A fwd comprising the first region (Fig. 5, PCR II); and (ii) a fourth oligonucleotide A rev, comprising the fourth region (Fig. 5, PCR II); (d) performing at least two cycles of PCR amplification, to generate a second PCR amplification product (2-4 cycles for second PCR in Fig. 5; paras. 0072-77)); (e) introducing to the second PCR amplification product: (iii) a seventh oligonucleotide BT/A2A, comprising the fourth region (Fig. 5, PCR III); and (f) performing at least two cycles of PCR amplification to generate a third PCR amplification product (multiple cycle in third PCR; paras. 0072-77). As to claim 2, WOOD teaches the first region in the first oligonucleotide in step (a) and the fourth region in the second oligonucleotide in step (a) generate binding sites for universal amplification performed in step (c) (Fig. 5). As to claim 3, WOOD teaches the fourth region in the second oligonucleotide comprises at least part of the next-generation sequencing (NGS) adapter sequence (Fig. 5). As to claim 4, WOOD teaches the melting temperatures of the first and the fourth regions are between 0.01° C. and 10° C. higher than the melting temperatures of the third and the seventh regions (compare paras. 0051 & 0056 with para. 0039 (target-specific can be 56°C versus universal can be 72°C)). As to claim 6, WOOD teaches the first PCR amplification product from step (d) is purified prior to step (e) using a method selected from the group consisting of SPRI purification, column purification, and enzymatic digestion (paras. 0031-32, 0068-69, 0072, 0079 & 0081). As to claim 8, WOOD teaches (g) introducing to the PCR amplification product obtained in step (f), (i) an eighth oligonucleotide, comprising from 5′ to 3′ end, a ninth region and an eighth region, wherein the ninth region comprises at least part of the next-generation sequencing (NGS) adapter sequence, and optionally (ii) a ninth oligonucleotide, comprising the fourth region (Fig. 5); and (h) performing at least one cycle of PCR amplification to obtain a third PCR amplification product (Fig. 5). As to claim 9, WOOD teaches (i) adding NGS adapter sequences to the PCR amplification product obtained in step (f) by ligation reaction (ligation as a known option; para. 0003). As to claim 14, WOOD teaches wherein an annealing temperature used in step (d) is between 0.01° C. and 10° C. higher than an annealing temperature used in step (b) (see claim 4; Examples, showing first anneal at 63°C and second at 64°C). As to claim 15, WOOD teaches at least one of the between 1 and 10,000 target genomic regions is selected from the group consisting of is selected from the group consisting of AKT1, ALK, APC, AR, ATM, BRAF, CCND1, CDK4, CDKN2A, CHEK2, CTNNB1, DDR2, EGFR, ERBB2, ERBB3, ERBB4, ESR1, EZH2, FBXW7, FGFR1, FGFR2, FGFR3, FGFR4, FLT3, FOXL2, GNA11, GNAQ, GNAS, HRAS, IDHL JAK1, JAK2, JAK3, KIT, KRAS, MAP2K1, MAP2K2, MET, MLH1, MPL, MTOR, MYC, MYCN, MYD88, NPM1, NRAS, PDGFRA, PIK3CA, PTEN, PTPN11, RAF1, RB1, RET, ROS1, SF3B1, SMAD4, SMARCB1, SMO, STK11, and TP53 (NRAS, EGFR, KRAS, para. 0120). WOOD does not teach (i) a fifth oligonucleotide (Blocker Displacement Amplification (BDA) forward primer) for each target genomic region, wherein the BDA forward primer comprises an eighth region targeting a specific genomic region, wherein the genomic region targeted by the eighth region is between 1 and 20 nucleotides closer to the seventh region compared to the genomic region targeted by the third region, (ii) a sixth oligonucleotide (BDA blocker) for each target genomic region, wherein 4 or more nucleotides at the 3′ end of the BDA forward primer sequence are also present at or near the 5′ end of the BDA blocker sequence; and wherein the BDA blocker contains a 3′ sequence or modification that prevents extension by a DNA polymerase, and wherein the concentration of the BDA blocker is at least 2 times that of the BDA forward primer. This, in summary, is a form of competitive blocker allele-specific PCR (CB-ASP), as explained in the specification. However, CB-ASP, including the same assay claimed, has been routine in the art for decades, regularly used to achieve greater SNP-detection sensitivity. For example, BIGLIA teaches CB-ASP that uses a 3’-blocked oligo to competitively hybridize with a primer, the 3’-end of the primer overlapping with the 5’-end of the 3’-blocked oligo (Fig. 1). In fact, “the blocking oligonucleotide is added at a concentration that is at least twice, preferably at least three times, at least 4 times or at least 5 times higher than the concentration of the primer oligonucleotide it competes with” (BIGLIA, para. 0049). Further, “[t]he overlap (i.e. the common sequence between the competitor primer and the blocking oligonucleotide) can comprise, or consist of, at least 30%, 40%, 50%, 60%, 70%, 80% or 85% of the blocking oligonucleotide”; “[o]r the overlap can comprise, or consist of, at least 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 nucleotides” (para. 0047). Finally, “[t]he amplification of the target sequence can be followed by analysis of the amplified sequence, e.g. by a method to precisely determine the mutation(s) of the target sequence” such that “[o]nce the amplification or enrichment of the target sequence is complete, the sample may thus be further processed, e.g., subjected to a sequencing reaction” such as “Single-molecule sequencing, second generation high throughput sequencing, [and] pyrosequencing . . .” (paras. 0074-75). All of this increases sensitivity, as is known for decades (Example, 2. Sensitivity Test Results Using Oligo Blocks According to the Invention; see also US20140017685, para. 0099 (“One important factor affecting the sensitivity of detecting mutations which are present at a low frequency in a sample is the concentration ratio between the first primer and the blocking oligonucleotide probe. Generally, the concentration ratio of first primer/labelled oligonucleotide probe is less than one”) and US20170233821, Abstract & para. 0028 (“An ultra-sensitive, specific methodology for detecting PIK3CA mutations . . . presence of corresponding competitive blocking unlabeled probes for each exon can avoid non-specific amplification of wild-type PIK3CA sequence increasing the sensitivity and the specificity of method”; “This unlabeled blocking probe is used for competitive blocking of the wild type allele and is added at a higher concentration than the mutant allele specific primer, e.g. 5 to 20 times or 10 times higher concentration of the allele specific primer. . . . Thus, the unlabeled blocking probe competes with the allele specific primer for increased sensitivity”)). In fact, Applicants have known this well before their effective filing (see US20170067090, Abstract & Figures, disclosing competitive blocker allele-specific PCR using the same primer-blocker parameters claimed here). In other words, a skilled artisan would have been familiar with the routine use of CB-ASP using higher-concentration competitive blocker to achieve greater SNP sequencing detection from greater amplification sensitivity and specificity. In sum, the prior art demonstrates conclusively that a skilled artisan would have been motivated to apply higher-concentration CB-ASP blockers to the SNP sequencing detection of the prior art such as WOOD to similarly achieve greater sequencing sensitivity and specificity with a reasonable expectation of success. Double Patenting- Obvious Type The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory obviousness-type double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); and In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969). A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on a nonstatutory double patenting ground provided the conflicting application or patent either is shown to be commonly owned with this application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. Effective January 1, 1994, a registered attorney or agent of record may sign a terminal disclaimer. A terminal disclaimer signed by the assignee must fully comply with 37 CFR 3.73(b). Instant claims 1-4, 6, 8, 9 and 14-15 are rejected on the ground of nonstatutory obviousness-type double patenting as being unpatentable over conflicting claims 1, 3-5, 9, 17, 22, 24-27, 29, 34-36, 40-41, 43, 47 and 52 of 17/420476, in view of BIGLIA (US20160076089) as evidenced by US20140017685 and US20170233821. The instant claims are obvious over the conflicting claims because the conflicting claims teach the same three-PCR sequencing library preparation as the conflicting claims, and adding a CB-ASP reaction was a familiar option with familiar results. More specifically, the conflicting claims teach: 1. A method for preparing targeted regions of genomic DNA for high-throughput sequencing, the method comprising: (a) obtaining a genomic DNA sample; (b) amplifying at least a portion of the genomic DNA sample by performing two cycles of PCR using: (i) a first oligonucleotide comprising, from 5′ to 3′, a first region, a second region having a length between 0 and 50 nucleotides, a third region comprising at least four degenerate nucleotides, and a fourth region comprising a sequence that is complementary to a first target genomic DNA region; and (ii) a second oligonucleotide comprising, from 5′ to 3′, a fifth region, a sixth region having a length between 0 and 50 nucleotides, and a seventh region comprising a sequence that is complementary to a second target genomic DNA region; (c) amplifying a product of step (b) by performing at least three cycles of PCR with an annealing temperature that is 0-10° C. higher than an annealing temperature used in step (b) and using: (i) a third oligonucleotide comprising a sequence that is able to hybridize to the reverse complement of at least a portion of the first region; and (ii) a fourth oligonucleotide comprising a sequence that is able to hybridize to the reverse complement of at least a portion of the fifth region; and (d) amplifying a product of step (c) by performing at least one cycle of PCR using a fifth oligonucleotide comprising, from 5′ to 3′, an eighth region, a ninth region having a length between 0 and 50 nucleotides, and a tenth region comprising a sequence that is complementary to a third target genomic DNA region, wherein the third target genomic DNA region is at least one nucleotide closer to the first target genomic DNA region than the second target genomic DNA region; wherein the resulting amplicons are suitable for quantitative next-generation sequencing to enable copy number variation (CNV) and allele ratio quantitation. The conflicting claims do not explicitly teach (i) a fifth oligonucleotide (Blocker Displacement Amplification (BDA) forward primer) for each target genomic region, wherein the BDA forward primer comprises an eighth region targeting a specific genomic region, wherein the genomic region targeted by the eighth region is between 1 and 20 nucleotides closer to the seventh region compared to the genomic region targeted by the third region, (ii) a sixth oligonucleotide (BDA blocker) for each target genomic region, wherein 4 or more nucleotides at the 3′ end of the BDA forward primer sequence are also present at or near the 5′ end of the BDA blocker sequence; and wherein the BDA blocker contains a 3′ sequence or modification that prevents extension by a DNA polymerase, and wherein the concentration of the BDA blocker is at least 2 times that of the BDA forward primer. This, in summary, is a form of competitive blocker allele-specific PCR (CB-ASP), as explained in the specification. However, CB-ASP, including the same assay claimed, has been routine in the art for decades, regularly used to achieve greater SNP-detection sensitivity. For example, BIGLIA teaches CB-ASP that uses a 3’-blocked oligo to competitively hybridize with a primer, the 3’-end of the primer overlapping with the 5’-end of the 3’-blocked oligo (Fig. 1). In fact, “the blocking oligonucleotide is added at a concentration that is at least twice, preferably at least three times, at least 4 times or at least 5 times higher than the concentration of the primer oligonucleotide it competes with” (BIGLIA, para. 0049). Further, “[t]he overlap (i.e. the common sequence between the competitor primer and the blocking oligonucleotide) can comprise, or consist of, at least 30%, 40%, 50%, 60%, 70%, 80% or 85% of the blocking oligonucleotide”; “[o]r the overlap can comprise, or consist of, at least 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 nucleotides” (para. 0047). Finally, “[t]he amplification of the target sequence can be followed by analysis of the amplified sequence, e.g. by a method to precisely determine the mutation(s) of the target sequence” such that “[o]nce the amplification or enrichment of the target sequence is complete, the sample may thus be further processed, e.g., subjected to a sequencing reaction” such as “Single-molecule sequencing, second generation high throughput sequencing, [and] pyrosequencing . . .” (paras. 0074-75). All of this increases sensitivity, as is known for decades (Example, 2. Sensitivity Test Results Using Oligo Blocks According to the Invention; see also US20140017685, para. 0099 (“One important factor affecting the sensitivity of detecting mutations which are present at a low frequency in a sample is the concentration ratio between the first primer and the blocking oligonucleotide probe. Generally, the concentration ratio of first primer/labelled oligonucleotide probe is less than one”) and US20170233821, Abstract & para. 0028 (“An ultra-sensitive, specific methodology for detecting PIK3CA mutations . . . presence of corresponding competitive blocking unlabeled probes for each exon can avoid non-specific amplification of wild-type PIK3CA sequence increasing the sensitivity and the specificity of method”; “This unlabeled blocking probe is used for competitive blocking of the wild type allele and is added at a higher concentration than the mutant allele specific primer, e.g. 5 to 20 times or 10 times higher concentration of the allele specific primer. . . . Thus, the unlabeled blocking probe competes with the allele specific primer for increased sensitivity”)). In other words, a skilled artisan would have been familiar with the routine use of CB-ASP using higher-concentration competitive blocker to achieve greater SNP detection and amplification sensitivity and specificity. In sum, the prior art demonstrates conclusively that a skilled artisan would have been motivated to apply higher-concentration CB-ASP blockers to the SNP sequencing detection of the conflicting claims to similarly achieve greater sequencing sensitivity and specificity with a reasonable expectation of success. Thus, the conflicting claims in light of the familiar CB-ASP prior art render obvious the instant claims. Instant claims 1-4, 6, 8, 9 and 14-15 are rejected on the ground of nonstatutory obviousness-type double patenting as being unpatentable over conflicting claims 1-12 of US12331350, in view of WOOD (US20210180121). The instant claims are obvious over the conflicting claims because the conflicting claims teach the same CB-ASP reaction for sequencing library preparation, and three-PCR sequencing library preparation of the conflicting claims was a familiar option with familiar results. More specifically, the conflicting claims teach: 1. A method for simultaneously amplifying and detecting allelic variants at at least ten genetic loci, the method comprising: (a) mixing a sample comprising DNA with a DNA polymerase and a blocker displacement amplification (BDA) oligo set for each genetic locus, each BDA oligo set comprising (i) a BDA forward primer, (ii) a BDA blocker, and (iii) a BDA reverse primer, wherein at least four nucleotides at the 3′ end of each BDA forward primer sequence are also present at or near the 5′ end of its respective BDA blocker sequence, wherein each BDA blocker contains a 3′ sequence or modification that prevents extension by DNA polymerase, and wherein the concentration of each BDA blocker is at least twice that of its respective BDA forward primer; and (b) subjecting the mixture to at least four cycles of amplification, thereby producing amplicons; (c) performing next-generation sequencing (NGS) of the amplicons. 2. The method of claim 1, wherein the DNA polymerase has 3′ to 5′ exonuclease activity. 3. The method of claim 2, wherein each BDA blocker has a 3′ modification that prevents 3′ to 5′ exonuclease activity. 4. The method of claim 1, wherein the concentration of each BDA reverse primer and/or each BDA forward primer is determined based on a reads analysis of a previous calibration NGS experiment, wherein the concentration of each BDA reverse primer and/or each BDA forward primer is increased relative to the concentration used for the previous calibration NGS experiment. 7. The method of claim 1, wherein the BDA oligo set comprises at least 10 BDA oligo sets, each BDA oligo set comprising (i) a BDA forward primer, (ii) a BDA blocker, and (iii) a BDA reverse primer, wherein at least four nucleotides at the 3′ end of each BDA forward primer sequence are also present at or near the 5′ end of its corresponding BDA blocker sequence, wherein each BDA blocker contains a 3′ sequence or modification that prevents extension by DNA polymerase, and wherein the concentration of each BDA blocker is at least twice that of its corresponding BDA forward primer, wherein each BDA blocker is complementary to a genomic region bearing a single nucleotide polymorphism (SNP) in which the alternative allele has a population frequency of between 10% and 90%, and wherein each corresponding BDA forward primer is not complementary to the SNP locus. The conflicting claims do not explicitly teach the three-PCR technique of the conflicting claims. However, this same technique was already known in the art to yield “overlapping primer pairs in a single tube for contiguous coverage over target regions, while simultaneously preventing amplification of both primer dimers and undesirable mini-amplicons that result from overlapping primer pairs” (WOOD, Abstract). Specifically, as explained above, WOOD teaches a method for labeling and amplifying each strand of between 1 and 10,000 target genomic regions with an oligonucleotide barcode sequence by polymerase chain reaction (PCR), the method comprising: (a) introducing into a DNA sample comprising the between 1 and 10,000 target genomic regions, for each target genomic region (Fig. 5): (i) a first oligonucleotide F1/F2, comprising from 5′ to 3′ end, (A) a first region, (B) a second region with a length between 0 and 50 nucleotides, and (C) a third region targeting a first specific genomic region (Fig. 5, PCR I); and (ii) a second oligonucleotide R1/R2, comprising from 5′ to 3′ end, (A) a fourth region, (B) a fifth region with a length between 0 and 50 nucleotides, (C) a sixth region comprising a unique molecular identifier (UMI) comprising at least four degenerate nucleotides, and (D) a seventh region targeting a second specific genomic region (Fig. 5, PCR I); (b) performing at least two cycles of PCR amplification to generate a first PCR amplification product (2-3 cycles for first PCR in Fig. 5; paras. 0072-77); (c) introducing into the first PCR amplification product: (i) a third oligonucleotide A fwd comprising the first region (Fig. 5, PCR II); and (ii) a fourth oligonucleotide A rev, comprising the fourth region (Fig. 5, PCR II); (d) performing at least two cycles of PCR amplification, to generate a second PCR amplification product (2-4 cycles for second PCR in Fig. 5; paras. 0072-77)); (e) introducing to the second PCR amplification product: (iii) a seventh oligonucleotide BT/A2A, comprising the fourth region (Fig. 5, PCR III); and (f) performing at least two cycles of PCR amplification to generate a third PCR amplification product (multiple cycle in third PCR; paras. 0072-77) In sum, the prior art demonstrates that a skilled artisan would have been motivated to apply the three-PCR SNP loci sequencing library preparation technique of the prior art to the sequencing library SNP loci preparation technique of the conflicting claims to “overlapping primer pairs in a single tube for contiguous coverage over target regions, while simultaneously preventing amplification of both primer dimers and undesirable mini-amplicons that result from overlapping primer pairs” with a reasonable expectation of success. Thus, the conflicting claims in light of the familiar multi-PCR sequencing library preparation prior art render obvious the instant claims. Prior Art The following prior art demonstrates that SNP-loci sequencing library preparation using barcodes and multiple PCR steps was well-known before effective filing of this application: US20200407798. Conclusion No claims are allowed. Any inquiry concerning this communication or earlier communications from the examiner should be directed to Aaron Priest whose telephone number is (571)270-1095. The examiner can normally be reached 8am-6pm. Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice. If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Gary Benzion can be reached at (571) 272-0782. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300. Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000. /AARON A PRIEST/Primary Examiner, Art Unit 1681
Read full office action

Prosecution Timeline

Nov 01, 2022
Application Filed
Jan 12, 2026
Non-Final Rejection — §103, §112, §DP (current)

Precedent Cases

Applications granted by this same examiner with similar technology

Patent 12601008
Using Hairpin Formation To Identify DNA and RNA Sequences Having A Target Nucleic Acid Sequence
2y 5m to grant Granted Apr 14, 2026
Patent 12595509
SYNTHETIC NUCLEIC ACID SPIKE-INS
2y 5m to grant Granted Apr 07, 2026
Patent 12590328
TARGETED DEPLETION OF NON-TARGET LIBRARY MOLECULES USING POISON PRIMERS DURING TARGET CAPTURE OF NEXT-GENERATION SEQUENCING LIBRARIES
2y 5m to grant Granted Mar 31, 2026
Patent 12589396
DIGITAL TO BIOLOGICAL CONVERTER
2y 5m to grant Granted Mar 31, 2026
Patent 12590341
BRASSICA GAT EVENT AND COMPOSITIONS AND METHODS FOR THE IDENTIFICATION AND/OR DETECTION THEREOF
2y 5m to grant Granted Mar 31, 2026
Study what changed to get past this examiner. Based on 5 most recent grants.

AI Strategy Recommendation

Get an AI-powered prosecution strategy using examiner precedents, rejection analysis, and claim mapping.
Powered by AI — typically takes 5-10 seconds

Prosecution Projections

1-2
Expected OA Rounds
61%
Grant Probability
87%
With Interview (+26.0%)
3y 0m
Median Time to Grant
Low
PTA Risk
Based on 794 resolved cases by this examiner. Grant probability derived from career allow rate.

Sign in with your work email

Enter your email to receive a magic link. No password needed.

Personal email addresses (Gmail, Yahoo, etc.) are not accepted.

Free tier: 3 strategy analyses per month