Apr 16, 2026
Protocol for Cleaning and Preparing Insect Specimens Trapped into Adhesive (e.g., TAD, Tanglefoot, Pestik) Prior to Diagnostic Identification and DNA Extraction
- Axel David Gonzalez Murillo1,
- William Klingeman III1,
- John Moulton1
- 1University of Tennessee
- Axel Gonzalez Murillo
Protocol Citation: Axel David Gonzalez Murillo, William Klingeman III, John Moulton 2026. Protocol for Cleaning and Preparing Insect Specimens Trapped into Adhesive (e.g., TAD, Tanglefoot, Pestik) Prior to Diagnostic Identification and DNA Extraction. protocols.io https://dx.doi.org/10.17504/protocols.io.4r3l2dr6qg1y/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 08, 2026
Last Modified: April 16, 2026
Protocol Integer ID: 314719
Keywords: dna extraction, preparing insect, dna extraction this protocol document, cleaner specimen, enabled cleaner specimen, recovered dna, trapped insect, specimen, trapping adhesive, use of specimen, drying method, specimen morphology, arthropod trap, removal of adhesive, removal of the adhesive, dna, protocol for cleaning, adhesive reside, submerged specimen, higher volumes of recovered dna, adhesive, pestik, tanglefoot, loss of dna, reliable molecular analysis, allowed reliable molecular analysis
Funders Acknowledgements:
USDA NIFA
Grant ID: #7002511
Abstract
This protocol documents a refined methodology for cleaning and preparing insect
specimens recovered from arthropod traps that utilize commercially available adhesive
products (e.g., TAD, Tanglefoot, Pestik) prior to use of specimen for diagnostic
identification and DNA extraction. Removal of the adhesive is achieved via
immersion in Histo-Clear II‱ for 30–45 minutes and benefits form periodic
agitation of the submerged specimen, followed by detergent-based washing in
warm water to eliminate adhesive resides. Subsequent rinsing and controlled
drying methods preserve specimen morphology while minimizing loss of DNA that
may occur through binding with residues from the trapping adhesive. Thorough
removal of adhesive has yielded higher volumes of recovered DNA from specimens trapped
using TAD adhesive. The procedure has enabled cleaner specimens, preserved integumental
setae (hairs) and delicate antenna, and has allowed reliable molecular analyses
of field-trapped insects.
Materials
-Histo-Clear II‱
-Dawn Ultra Platinum
-Optional (Hairdryer, Computer dust cleaner)
-Light diffuser
Protocol step by step for Cleaning and Preparing Insect Specimens
Initial Glue Removal
Carefully remove specimen from adhesive trap surface; sun-exposed warm adhesive is generally more fluid, enabling retention of delicate appendages, setae, etc.
- Several different brands of insect adhesive are commercially available, including, but not limited to, TAD (Trécé Adhesive Division, Adair, OK, USA), Tanglefoot (The Tanglefoot Company, Grand Rapids, MI, USA), and Pestik (Phytotronics Inc., Earth City, MO, USA).
- Note: Lower than expected recovery of DNA from specimens that still had surface residues of adhesives that had been used during field trapping. It is likely that the DNA may become bound to the adhesive during the DNA extraction process (see additional information below).
- Most specimens successfully cleaned, from which DNA was successfully extracted, were those that had been recovered from TAD adhesive.
Submerge in Solvent Bath
Submerge in Solvent Bath: Place the beetle in a Histo-Clear II‱ (National Diagnostics, Atlanta, GA, USA) (low-hazard histological clearing agent) in a small glass vial or container. Allow the specimen(s) to be completely submerged for 30 to 45 minutes.
- It is usually not necessary to submerge specimens for longer than 60 minutes, provided that the specimen is regularly agitated.
- Open (air) space in the rest of the vial is okay and can allow for easier agitation of the solvent.
- Note: For fragile or rare/uncommon specimens, specimens (rather than as a grouping of multiple specimens) can be submerged independently during the adhesive removal step.
Agitation
While the specimen is submerged, gently tilt the container two to four times from a vertical to horizontal orientation at about five-minute intervals across the 30-to-45-minute soak time.
- This action helps to dissolve and remove the adhesive from the specimen body without causing physical damage to the specimen.
- Note: some adhesives may aggregate into clumps and irregular balls that can include other environmental particles and accumulate in crevices and integumental sutures.
Transfer
Once the targeted soak time has elapsed, remove the beetle and place it in a glass or plastic petri dish lined with low-lint, absorbent lab tissue paper or wipes. Slightly moist, still pliable specimens can be used in proceeding to the next step.
Pinning and Deep Cleaning
Proceed by properly mounting the specimen on an insect pin to facilitate easier handling of the sample during the following cleaning actions.
- Detergent Wash: Dip the pinned sample into a solution of Dawn Ultra Platinum (grease cleaning agent) (Procter & Gamble Co., Cincinnati, OH, USA) detergent and warm (approx. 100 oF/38 oC degree) water, as cold water hardens adhesive residue, making it difficult to remove. Hot water should be avoided because temperatures exceeding 266 oF/130 oC may initiate degradation of specimen DNA.
- Manual Cleaning: If adhesive residue is still visible, use a fine, synthetic bristle artist paintbrush to gently brush away aggregate particles and to re-establish orientation or loft of densely distributed setae (e.g., on the frons of the head).
- Rinse: Rinse the beetle thoroughly using a wash bottle filled with warm water.
Drying and Inspection
Drying: Dry the specimen using one of the following methods:
- Natural: Leave the pinnedspecimen to dry naturally at ambient lab temperature. It may be helpful to cover the specimen beneath a vented plastic cover to prevent the accumulation of airborne particulates, fibers, and dust from clothing and the surrounding area.
- Hairdryer: Use a dryer set to no/low heat and low airflow speed to avoid blasting the specimen with too much air, particularly when handling fragile specimens and those with long appendages and dense setae (hairs).
- Compressed Air: Use a pressurized can of air (e.g., computer dust cleaner)
1. Some beetle specimens (e.g., metallic woodboring beetles (Buprestidae), click beetles (Elateridae), etc. may be sturdy enough to withstand heavier air flow volumes that are applied directionally toward the head-end (Fig.1).
Figure 1. Drying a pin-mounted adult buprestid beetle specimen using the compressed
air method.
2. Pressurized air volume can be adjusted by using modified straws, hoses, pipette tips, a pen barrel, etc. to change the diameter of the air exit port (narrower diameter = faster airflow) (Fig.2).
Figure 2. Air speed from compressed air, or an electric computer dust cleaner can be modified
using a pen barrel supplemented with a plastic straw.
3. Evaluate: Inspect the specimen. If the specimen is not optimally clean, still appears to have adhesive residue, or is still coated in environmental detritus (sand, fibers, pollen, etc.), repeat the detergent, rinsing and drying steps.
(Supplemental and Optional)
A Resourced and Aid for Visualizing Topography and Structural Morphology on High Reflectance (e.g., metallicized) Exoskeletal Surfaces.
- Light Diffusion: Because metallic woodboring beetles (also called Jewel Beetles) often have metallicized or highly reflective external surfaces (including within foveae on the elytral surface), some structures, features, on topography on integumental surfaces may be seen more clearly by placing specimens within a fully circular or partial light-diffusing ring of opaque plastic (e.g., rings cut from of Nalgene bottle sides (Thermo Fisher Scientific, Rochester, NY, USA), or a sheet/ring of transparent (e.g., rice paper) paper that separates the exoskeletal surface from direct exposure to light from the illumination source Fig.3).
Figure 3. A beetle specimen under the microscope with a light diffuser using cut, opaque plastic rings made from the sides of 200 mL Nalgene bottle (a rice-paper and foam light diffuser is visible in the background).
Final ID
Proceed with the identification of the specimen under the scope.