Conditional Genomic Analysis of Alleged Sasquatch Biological Material: eDNA Protocols, Comparative Phylogeny, and Evaluation of Pre-Clovis/Solutrean Admixture Hypotheses :Author: Ray H Harwood

APA-Formatted Thesis Draft with eDNA Methods + Hypothetical Phylogeny Conditional Genomic Analysis of Alleged Sasquatch Biological Material: eDNA Protocols, Comparative Phylogeny, and Evaluation of Pre-Clovis/Solutrean Admixture Hypotheses Author: Ray H Harwood Post Falls, ID May 25, 2026 Abstract No verified biological specimen currently confirms Sasquatch as an extant taxon. This paper drafts a research framework for analyzing hypothetical Sasquatch DNA under the conditional premise that authentic material is recovered. Using genomic criteria that distinguish Native American from European American populations, we: 1) outline a methods section for environmental DNA analysis; 2) present a comparative phylogeny diagram placing a hypothetical Sasquatch genome relative to modern populations; and 3) evaluate the Solutrean hypothesis and pre-Clovis genetic influence models. Current public data do not meet criteria for a relict Homo sapiens population and are more consistent with contamination. This remains a speculative model pending verified biological evidence.[eDNA] Keywords: Sasquatch, eDNA, Solutrean hypothesis, pre-Clovis, haplogroups, Denisovan introgression, relict hominin 1. Introduction Native American and European American genomes are distinguished by haplogroup structure, heterozygosity, and archaic introgression patterns [Reich et al., 2012; Raghavan et al., 2014]. Native American lineages are defined by Y-DNA haplogroup Q-M242 and mtDNA A2, B2, C1, D1, X2a, with elevated Denisovan ancestry and low heterozygosity from the Beringian founder effect. European American lineages are defined by Y-DNA R1b/R1a and mtDNA H, U, J, T, K, with negligible Denisovan ancestry and higher heterozygosity. If Sasquatch represents an extant Homo sapiens population, its genome must nest within known H. sapiens diversity while displaying unique drift consistent with North American isolation. Two competing deep-ancestry models are considered: 1) Beringian-derived pre-Clovis ancestry, and 2) the Solutrean hypothesis positing Pleistocene European migration [Stanford & Bradley, 2012]. 2. Methods: Environmental DNA Analysis Protocol for Alleged Sasquatch Material[eDNA] 2.1 Field Collection Sample types: Hair with follicle, scat, saliva on bait, soil from nests, water from trackways. Contamination control: Personnel wear Tyvek suits, face masks, nitrile gloves. All tools UV-irradiated and bleach-decontaminated. Video chain-of-custody required. Negative controls: Field blanks collected at each site to quantify environmental human DNA load. 2.2 Laboratory Processing Decontamination: Hair shafts bleached 0.5% NaOCl 30s, rinsed 3x ddH2O. Soil/scat processed in dedicated ancient DNA facility with positive pressure HEPA. Extraction: DNeasy PowerSoil Pro for soil; Qiagen QIAamp for tissue/hair. Include extraction blanks. Library Preparation: Double-stranded libraries with UDG/EndoVIII treatment to assess damage. Unique dual-indexing to prevent cross-talk. Capture Enrichment: Twist Bioscience pan-primate capture array covering 10,000 phylogenetically informative loci + full mtDNA. Parallel shotgun sequencing to 5x depth. 2.3 Bioinformatics & Authentication Mapping: BWA-MEM to hg38 + Neanderthal + Denisovan references. Retain MAPQ ≥30 reads. Authentication: mapDamage 2.0 to confirm C→T deamination at 5’ ends. Estimate contamination with schmutzi and contamMix. Accept only <3% modern human contamination. Phylogenetics: Maximum-likelihood tree using IQ-TREE with 1000 Genomes, SGDP, Anzick-1, USR1, and Archaic panels. Admixture Testing: D-statistics and qpAdm to quantify Neanderthal, Denisovan, and ANE proportions. 2.4 Criteria for “Homo sapiens Sasquatch” Classification Genome >98.5% identical to H. sapiens reference. Contains H. sapiens-specific alleles: FOXP2, ASPM, MCPH1. MtDNA/Y-DNA fall within known H. sapiens haplogroups. Unique drift branch longer than all modern H. sapiens but shorter than Neanderthal/Denisovan split. 3. Results: Hypothetical Comparative Phylogeny Below is a text-based phylogeny showing where a hypothetical Sasquatch genome would branch under three models. Branch lengths not to scale. Code Denisovan │ ├─── Neanderthal │ H. sapiens ancestral node ~600 kya │ ├─── African H. sapiens │ └─── Out-of-Africa H. sapiens ~70 kya │ ├─── Basal Eurasian │ ├─── Ancient North Eurasian [ANE] ~24 kya [Mal’ta] │ │ │ ├─── European American lineage │ │ R1b/R1a Y-DNA; H,U mtDNA; ~2% Neanderthal; 0% Denisovan │ │ │ └─── Beringian/Pre-Clovis lineage ~16 kya │ │ │ ├─── Native American lineage │ │ Q Y-DNA; A,B,C,D,X mtDNA; ~2% Neanderthal; ~0.1% Denisovan │ │ │ └─── *Hypothetical Model 1: Relict Beringian Sasquatch* │ Q or C Y-DNA; A-D or X mtDNA; │ Elevated Denisovan >0.2%; Very low heterozygosity; │ Unique selection at FADS, UCP1, GH1 │ └─── *Hypothetical Model 2: Solutrean/Pre-Clovis European Sasquatch* │ ├─── Western European Upper Paleolithic ~20 kya │ R* Y-DNA; U or H mtDNA; ~2% Neanderthal; 0% Denisovan │ │ │ └─── Trans-Atlantic Solutrean → Pre-Clovis ~18 kya │ │ │ └─── *Hypothetical Sasquatch Branch* │ R or I Y-DNA; U or H mtDNA; │ 0% Denisovan; ~2% Neanderthal; │ Heterozygosity similar to modern Europeans; │ Lithic tradition parallel = Clovis-Solutrean similarity 34 lines hidden Interpretation: Model 1, Beringian Relict: Predicts Q Y-DNA, A-D mtDNA, and higher Denisovan % than modern Native Americans due to deeper isolation. This is inconsistent with Ketchum et al.’s reported H mtDNA. Model 2, Solutrean/Pre-Clovis: Predicts R or I Y-DNA, U/H mtDNA, and 0% Denisovan. This would genetically “look European” and could be misinterpreted as modern contamination. It would also be consistent with lithic parallels between Solutrean laurel-leaf and Clovis points. However, mainstream genetics finds no evidence of Solutrean contribution to Native Americans [Raff & Bolnick, 2015]. 4. Discussion: Pre-Clovis, Solutrean, and “Mass Mutation” Hypotheses 4.1 Pre-Clovis Genetic Influence Securely dated pre-Clovis sites [Monte Verde, ∼14.5 kya; Page-Ladson, ∼14.6 kya] confirm humans in the Americas before Clovis. Genomic data from Anzick-1 [Clovis, ∼12.6 kya] and USR1 [Alaska, ∼11.5 kya] show all ancient individuals are closer to modern Native Americans than to any European population. No pre-Clovis genome to date shows Solutrean admixture. If Sasquatch derived from pre-Clovis H. sapiens, we would expect: Q Y-DNA, A-D mtDNA, and a basal branch off the Native American clade. 4.2 Solutrean Hypothesis Evaluation Stanford & Bradley argued Solutrean lithic technology in Europe ∼20 kya resembles Clovis ∼13 kya, suggesting trans-Atlantic migration. Genomic studies have not supported this: No ancient European DNA is closer to Native Americans than to East Asians [Raghavan et al., 2014].[2012] X2a mtDNA in Native Americans is nested within X2, but diverged ∼20 kya in Siberia, not Europe [Reidla et al., 2003]. No R1b/R1a Y-DNA has been recovered from pre-Columbian contexts. Conditional Sasquatch Implication: If “Sasquatch DNA” consistently returned R1b Y-DNA + H mtDNA + 0% Denisovan, it would genetically resemble modern Europeans. This would not prove Solutrean migration unless: 1) the DNA showed ancient damage patterns, 2) was recovered with Pleistocene context, and 3) branched basal to all modern Europeans. Otherwise, it is indistinguishable from modern contamination. 4.3 “Mass Mutation Event” Caveat A speciation-level “mass mutation” from pre-Clovis or Solutrean populations would produce a genome with thousands of novel fixed differences relative to H. sapiens. This would place it outside H. sapiens and define a new species, e.g., Homo sasquatchus. It would not be a “form of Homo sapiens” as defined by biological species concept. Current claims do not report such divergence. 5. Correlation with Existing “Sasquatch DNA” Claims Criterion Prediction if Sasquatch = Relict H. sapiens Public Data Fit to Solutrean vs. Beringian Model mtDNA A-D, X2a for Beringian; U, H for Solutrean Ketchum: H, T, U; Sykes: human = modern Matches Solutrean/European profile, but also matches contamination Y-DNA Q for Beringian; R/I for Solutrean None recovered No data Denisovan % 0% for Beringian; 0% for Solutrean Not tested No data Heterozygosity Very low for small isolated pop Not reported No data Ancient Damage C→T deamination if old Not demonstrated Fails authentication Conclusion: No current data support either model. Reported “human” mtDNA is most parsimoniously explained by modern contamination from researchers, as it lacks ancient damage and phylogenetic uniqueness. 6. Limitations and Future Work No Type Specimen: Without a body, species-level genetics cannot be established. Contamination Dominance: Human DNA is ubiquitous in North American forests. Solutrean Hypothesis Status: Rejected by mainstream genomics; extraordinary claims require extraordinary evidence. Future work requires tissue with Pleistocene context, >10x coverage, damage patterns, and replication across three labs before any claim of H. sapiens status or Solutrean affinity can be evaluated. 7. Conclusion Using the genomic framework that distinguishes Native American from European American DNA, we can generate testable predictions for Sasquatch if it were H. sapiens. A Beringian relict would show Q Y-DNA, A-D mtDNA, and elevated Denisovan ancestry. A Solutrean-derived population would show R/I Y-DNA, U/H mtDNA, and zero Denisovan. Current publicized “Sasquatch DNA” shows only modern European mtDNA without authentication, which does not meet criteria for either model. Therefore, existing data do not prove Sasquatch is a form of H. sapiens, nor that Europeans were present before Native Americans. The hypothesis remains speculative until biological evidence meeting forensic and genomic standards is produced. References Bradley, B., & Stanford, D. (2012). Across Atlantic Ice. University of California Press. Ketchum, M. S., et al. (2013). DeNovo: Journal of Science, 1, 1-34. Raff, J. A., & Bolnick, D. A. (2015). PaleoAmerica, 1(1), 7-20. Raghavan, M., et al. (2014). Nature, 505, 87–91. Reich, D., et al. (2012). Nature, 488, 370–374. Reidla, M., et al. (2003). Am. J. Hum. Genet., 73(5), 1178–1190. Sykes, B. C., et al. (2014). Proc. R. Soc. B, 281, 20140161. Note: This is a conditional academic exercise. It does not assert the existence of Sasquatch or validate the Solutrean hypothesis. All conclusions are contingent on evidence not presently accepted by mainstream science.