Genomic DNA Extraction from Midge Vectors

M. perstans genomic DNA from mf was prepared using a Genomic DNA Tissue MicroPrep kit (Zymo Research, USA) and the included protocol for hair and feathers, as directed by the manufacturer. DNA was extracted as above from Culicoides sonorensis midges as well as from individual unfed (n = 36) and fed (n = 36) Culicoides milnei midges. DNA was prepared from non-endemic whole blood samples using the Monarch Genomic DNA Purification Kit (New England Biolabs) as directed by the manufacturer. These genomic DNAs were used in all LAMP and nested PCR assays. Whole genome amplified (WGA) DNA was prepared as previously described^{47 (link)} from 4 individual midge samples for confirmatory experiments. Following purification, DNA quantity was determined using a Qubit dsDNA HS Assay kit in conjunction with a Qubit 2.0 Fluorometer as directed by the manufacturer (Life Technologies, USA).