Development of Microsatellite Markers for Loxosceles devia Gertsch & Mulaik (Araneae: Sicariidae) using Next Generation Sequencing
Garza, Tanya Y
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Brown recluse spiders in the genus Loxosceles are known for causing severe necrotic skin lesions. Despite their impact on human health, very little is known concerning the genetic diversity or population structure of this genus. The development of a suite of molecular markers (microsatellites) would allow us to obtain genetic data and expand the resources available to study Loxosceles devia, the species found throughout south Texas. Microsatellites are long tandem repeats about 1-6 nucleotides long of DNA, also known as simple sequence repeats. Their co-dominant method of inheritance, wide spread distribution across the nuclear genome, ease of scoring, and high mutation rates make microsatellites an ideal molecular marker. Unfortunately, the cost of developing microsatellites has limited their availability until now. The Ion Torrent Personal Genome Machine® was used to shotgun sequence libraries of genomic DNA derived from L. devia. The program MSATCOMMANDER™ was used to identify and design primers for di-, tri-, and tetra- microsatellite repeats from these shotgun sequences. Seventeen microsatellite markers were designed and polymerase chain reaction conditions were optimized for each primer to develop a suite of novel molecular markers for this species.