title: Raccoon_USA_N2
design: DNA was extracted from skin and muscle tissues using NucleoSpin Tissue and NucleoSpin Tissue XS kit for DNA from cells and tissue (Macherey-Nagel) according to the manufacturer's protocol in the pre-PCR dedicated area. DNA was quantified using a Qubit Fluometer (ThermoFisher Scientific) and subsequently diluted to a final concentration of 20 ng/ul. Restriction site-Associated DNA (RAD) sequencing libraries were constructed following the Adapterama III protocol (Bayona-Vsquez et al., 2019) using PstI-HF, DdeI, and NsiI enzymes (New England Biolabs) together with relevant adapters indexed in accordance with design 3 from the aforementioned protocol. After digestion and ligation and prior to purification and amplification, all samples were pooled in equal volumes. At each nucleic acid purification step, short fragments and unincorporated adapters were removed using MagSi-NGS PREP Plus beads (magtivio) with a left-side selection procedure. After 12 cycles of PCR, libraries were visually verified via gel electrophoresis on a 1.5% agarose gel and quantified using a Qubit Fluometer. Size selection was performed using Pippin Prep (Sage Science) with the range of 400-600 bp