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26 December 2018 Method for DNA Isolation From Sweetpotato Weevil (Coleoptera: Curculionidae) Collected in Pheromone-Baited Traps
Phillip A. Wadl, Livy H. Williams, Karen R. Harris-Shultz, Grant T. McQuate
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Abstract

This study provides a protocol for the isolation of high-quality DNA from sweetpotato weevils (Cylas formicarius elegantulus (Summers)) collected from pheromone-baited aerial funnel traps.This study was based on our discovery that a 2-wk collection interval of sweetpotato weevils from pheromone traps did not permit isolation of intact high-quality genomic DNA. To test the effect of collection methods, i.e., sample collection interval and preservation method, on quality of isolated DNA, we placed freshly killed male sweetpotato weevils into aerial funnel traps in the field and removed subsamples at several times thereafter. DNA yield from freshly isolated (day = 0) samples was significantly greater than samples preserved in 70% ethanol or at -20°C, whereas there was no difference between 70% ethanol and -20°C storage. Likewise, DNA yield from freshly isolated (day = 0) samples was significantly greater than for later sampling times. Quality assessment of genomic DNA through gel electrophoresis and polymerase chain reaction (PCR) indicated isolation of high molecular weight DNA for all samples collected at t ≤ 7 d, but that DNA quality was degraded by 14 d. Our goal was to develop a reliable method for isolation of genomic DNA from field-collected sweetpotato weevil suitable for direct use in PCR. We discovered that it is critical to collect specimens from traps at an interval of 1 wk or less. Our findings allow for scheduling of sampling at reasonable intervals without the need for special materials.This has the added benefit of allowing individuals without special training to collect and prepare sweetpotato weevil specimens for genetic studies.

Published by Oxford University Press on behalf of Entomological Society of America 2018. This work is written by (a) US Government employee(s) and is in the public domain in the US.
Phillip A. Wadl, Livy H. Williams, Karen R. Harris-Shultz, and Grant T. McQuate "Method for DNA Isolation From Sweetpotato Weevil (Coleoptera: Curculionidae) Collected in Pheromone-Baited Traps," Journal of Economic Entomology 112(2), 1001-1003, (26 December 2018). https://doi.org/10.1093/jee/toy390
Received: 30 August 2018; Accepted: 19 November 2018; Published: 26 December 2018
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KEYWORDS
aerial funnel trap
Cylas formicarius elegantulus (Summers)
DNA isolation
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