Supplementary Information from High-quality fossil dates support a synchronous, Late Holocene extinction of devils and thylacines in mainland Australia

Detailed explanation of our statistical analyses, including SI Figure 1 and SI Tables 2-

Histological appearance of buried teeth at 40x (A, B) and 200x (C) magnification after A: one month—pulp tissue shows loss of structure and nuclear material; B:16 months—very little pulp tissue remains with much of the pulp chamber being empty.

<p>Dentine displayes almost complete loss of the predentine layer; and C: 16 months—cementum shows little structural change but loss of cells in soft tissue inclusions and on the external surfaces.</p

Histological appearance of teeth with a PMI >500 years at 40x (A) and 100x (B) magnification.

<p>A: Marked structural breakdown of the dentine and no visible cellular material. Areas of focal destruction in the cementum; and B: Destruction of the dentine traveling outwards along the dentinal tubules with almost the full thickness of the dentine being affected.</p

Predicted fragment yield for the mtDNA 77 bp fragment at varying PMI and soil temperature.

<p>PMI and soil temperature are held constant at sample median values where applicable.</p

Details of primers used for quantification of DNA.

<p>Details of primers used for quantification of DNA.</p

STR genotyping success for three dental tissues from five post-mortem burial intervals.

<p>Nineteen teeth were sampled for each time period (1, 2, 4, 8 and 16 months). Allele and complete locus dropout are expressed with colder colours representing lower success and warmer colours representing higher success.</p

Predicted DNA half-life for varying average soil temperature for each fragment type.

<p>For nuclear DNA as the half-life is affected by donor age the average donor age has been selected for the calculation.</p

DNA yields at each PMI interval (1–16 months) for three tooth tissues (cementum, coronal dentine and root dentine) for two nuclear DNA targets (67 bp and 156 bp) and one mtDNA target (77 bp).

<p>A-C: DNA yield for each tooth, and D-F: Arithmetic mean DNA yield at each PMI interval.</p

Predicted fragment yield for nuclear targets at varying PMI, soil temperature and subject age.

<p>PMI, soil temperature and subject age are held constant at sample median values where applicable. (Note the effect of PMI was much greater than that of soil temperature and age, the Y-axis scales reflect this.)</p

Genotype data

Geneotype data from samples collected along the Glenburn transect. Field number - individual, Site number - corresponds with Figure 1, Cline distance - distance along the transect and used in cline analysis, mtDNA - mtDNA identity as determined from RFLP, MS - Le=L. ewingii, Lp=L. paraewingi, Hyb=hybrid as determined from Structure analysis, YrCollected - year sample was collected, Le1-Le8 - genotype data from microsatellite loci