Dna extraction

DNA extraction

Specimens were extracted following a nondestructive procedure ( Beati et al., 2012; Beati & Keirans, 2001) where a small cut is made in the posterolateral abdomen before placing the tick in 180 µl Buffer ATL and 20 µl Proteinase K (Qiagen Inc., Valencia, CA) and incubating overnight at 56°C. After incubation, the exoskeleton is removed and returned to an ethanol-filled cryovial for preservation. The remainder of the extraction follows the manufacturer’s protocol for the Qiagen DNeasy Blood & Tissue Kit except for the elution step, where two successive elutions with 25 µl hot (72°C) Buffer AE are performed in the same tube for a final elution volume of 50 µl (30 µl for larvae).