TY - JOUR

T1 - Gauging DNA degradation among common insect trap preservatives

AU - Ruppert, Laura Sophia

AU - Segelbacher, Gernot

AU - Staab, Michael

AU - Winiger, Nathalie

N1 - Publisher Copyright: © 2023 The Authors. Entomologia Experimentalis et Applicata published by John Wiley & Sons Ltd on behalf of Netherlands Entomological Society.

PY - 2023/3

Y1 - 2023/3

N2 - Genetic methods for species identification are becoming increasingly popular and can accelerate insect monitoring. However, obtaining good DNA quality and quantity from insect traps remains a challenge for field studies. Ethylene glycol, propylene glycol, and Renner solution have been previously suggested as suitable preservatives for the collection of genetic material, but a systematic overview of their performance under compromising field conditions is lacking. Here we experimentally test whether and how different preservatives affect DNA quality under different conditions and evaluate how choice of preservative may affect metabarcoding and more demanding downstream applications (e.g., RADseq). For this, we used the house cricket, Acheta domesticus (L.) (Orthoptera: Gryllidae), and tested propylene glycol, ethylene glycol, and Renner solution for their ability to preserve DNA over 27 days in various dilutions and temperatures. DNA quality was measured as DNA fragmentation and success rates in PCR amplifying a COI fragment of 658, 313, or 157 bp. Undiluted propylene glycol and ethylene glycol always retained high molecular weight DNA at room temperature. No high molecular weight DNA was preserved at 37 °C or in any dilution. Nevertheless, the COI sequence could be amplified from samples at every condition. Renner solution did not preserve high molecular weight DNA and fragmentation increased over time at 37 °C until amplification was impossible. The results suggest that propylene glycol and ethylene glycol are suitable preservatives for collecting both genetic and morphological material, but dilution or high temperatures compromise their ability to preserve high molecular weight DNA. For genomic approaches requiring high DNA quality, additional preservatives may need to be tested.

AB - Genetic methods for species identification are becoming increasingly popular and can accelerate insect monitoring. However, obtaining good DNA quality and quantity from insect traps remains a challenge for field studies. Ethylene glycol, propylene glycol, and Renner solution have been previously suggested as suitable preservatives for the collection of genetic material, but a systematic overview of their performance under compromising field conditions is lacking. Here we experimentally test whether and how different preservatives affect DNA quality under different conditions and evaluate how choice of preservative may affect metabarcoding and more demanding downstream applications (e.g., RADseq). For this, we used the house cricket, Acheta domesticus (L.) (Orthoptera: Gryllidae), and tested propylene glycol, ethylene glycol, and Renner solution for their ability to preserve DNA over 27 days in various dilutions and temperatures. DNA quality was measured as DNA fragmentation and success rates in PCR amplifying a COI fragment of 658, 313, or 157 bp. Undiluted propylene glycol and ethylene glycol always retained high molecular weight DNA at room temperature. No high molecular weight DNA was preserved at 37 °C or in any dilution. Nevertheless, the COI sequence could be amplified from samples at every condition. Renner solution did not preserve high molecular weight DNA and fragmentation increased over time at 37 °C until amplification was impossible. The results suggest that propylene glycol and ethylene glycol are suitable preservatives for collecting both genetic and morphological material, but dilution or high temperatures compromise their ability to preserve high molecular weight DNA. For genomic approaches requiring high DNA quality, additional preservatives may need to be tested.

KW - Acheta domesticus

KW - barcoding

KW - DNA fragmentation

KW - DNA quality

KW - ethylene glycol

KW - Gryllidae

KW - metabarcoding

KW - monitoring

KW - Orthoptera

KW - propylene glycol

KW - Renner solution

KW - species identification

KW - Biology

KW - Educational science

UR - http://www.scopus.com/inward/record.url?scp=85146079751&partnerID=8YFLogxK

U2 - 10.1111/eea.13266

DO - 10.1111/eea.13266

M3 - Journal articles

AN - SCOPUS:85146079751

VL - 171

SP - 218

EP - 226

JO - Entomologia Experimentalis et Applicata

JF - Entomologia Experimentalis et Applicata

SN - 0013-8703

IS - 3

ER -