Sample collection and DNA/RNA extraction
A single adult male Tasmanian scrubtit was captured using a mist net at
Weilangta Forest in south-eastern Tasmania and transported immediately
to Hobart where it was euthanized by a veterinary surgeon (Australian
National University Animal Research Authority A2021/33). The sample was
dissected at the Tasmanian Museum and Art Gallery specimens collection,
where brain and organ tissue were preserved in RNALater before being
stored at -80˚C. High molecular weight (HMW) DNA was extracted from
heart and kidney tissue using the Nanobind Tissue Big DNA Kit v1.0 11/19
(Circulomics). A Qubit fluorometer was used to assess the concentration
of DNA with the Qubit dsDNA BR assay kit (Thermo Fisher Scientific). RNA
was extracted from six organs preserved in RNALater, using the RNeasy
Plus Mini Kit (Qiagen) with RNAse-free DNAse (Qiagen) digestion.
Extractions were performed using tissue from the heart, brain, spleen,
kidney, liver and gonads.