DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures

Autores
Stazione, Leonardo; Lantschner, María Victoria; Corley, Juan Carlos; Soliani, Carolina
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Molecular tools are increasingly used in entomology for several applications such as taxonomy, genetics, ecology and evolution. For these studies, DNA extraction from individual insects is a crucial step, since the yield and quality vary depending on the methods used. Finding an ideal balance between quality and yield is particularly difficult to reach and several constraints must be considered upon selecting the final protocol. Bark beetles (Coleoptera: Curculionidae, Scolytinae) are a diverse group of forest insects for which molecular studies at the individual level are important, given that many species are invasive and may become pests. However, DNA extraction in bark beetles is not easy, given their small size and complex molecules of diverse nature conforming their exoskeleton and present in their digestive tract. Here, we carried out a comparative analysis of DNA extraction performance in five bark beetle species: Hylurgus ligniperda, Hylastes ater, Orthotomicus erosus, Orthotomicus laricis and Cyrtogenius luteus. By assessing the efficiency of two different protocols, our aim was to establish the best species-specific method for population-level studies. Our results showed that a method’s whole performance mainly depends on the species considered and translates into DNA quantity and quality variance. We also noted that the traditional method showed better PCR efficiency for the smallest species whereas the commercial kit performed better for the larger beetles. Our comparative analysis provides evidence that no single method for DNA isolation is best, and that each particular species requires optimization.
Fil: Stazione, Leonardo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina. Università degli Studi di Firenze; Italia
Fil: Lantschner, María Victoria. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina
Fil: Corley, Juan Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina
Fil: Soliani, Carolina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina
Materia
DNA ISOLATION
SCOLYTINAE
DNA QUALITY AND QUANTITY
INSECT SIZE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/265119
Acceder
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network_name_str CONICET Digital (CONICET)
spelling DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular proceduresStazione, LeonardoLantschner, María VictoriaCorley, Juan CarlosSoliani, CarolinaDNA ISOLATIONSCOLYTINAEDNA QUALITY AND QUANTITYINSECT SIZEhttps://purl.org/becyt/ford/4.1https://purl.org/becyt/ford/4Molecular tools are increasingly used in entomology for several applications such as taxonomy, genetics, ecology and evolution. For these studies, DNA extraction from individual insects is a crucial step, since the yield and quality vary depending on the methods used. Finding an ideal balance between quality and yield is particularly difficult to reach and several constraints must be considered upon selecting the final protocol. Bark beetles (Coleoptera: Curculionidae, Scolytinae) are a diverse group of forest insects for which molecular studies at the individual level are important, given that many species are invasive and may become pests. However, DNA extraction in bark beetles is not easy, given their small size and complex molecules of diverse nature conforming their exoskeleton and present in their digestive tract. Here, we carried out a comparative analysis of DNA extraction performance in five bark beetle species: Hylurgus ligniperda, Hylastes ater, Orthotomicus erosus, Orthotomicus laricis and Cyrtogenius luteus. By assessing the efficiency of two different protocols, our aim was to establish the best species-specific method for population-level studies. Our results showed that a method’s whole performance mainly depends on the species considered and translates into DNA quantity and quality variance. We also noted that the traditional method showed better PCR efficiency for the smallest species whereas the commercial kit performed better for the larger beetles. Our comparative analysis provides evidence that no single method for DNA isolation is best, and that each particular species requires optimization.Fil: Stazione, Leonardo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina. Università degli Studi di Firenze; ItaliaFil: Lantschner, María Victoria. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; ArgentinaFil: Corley, Juan Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; ArgentinaFil: Soliani, Carolina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; ArgentinaCold Spring Harbor Laboratory Press2024-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/265119Stazione, Leonardo; Lantschner, María Victoria; Corley, Juan Carlos; Soliani, Carolina; DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures; Cold Spring Harbor Laboratory Press; bioRxiv; 4-2024; 1-262692-8205CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2024.04.06.588396v1.article-infoinfo:eu-repo/semantics/altIdentifier/doi/10.1101/2024.04.06.588396info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:37:53Zoai:ri.conicet.gov.ar:11336/265119instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:37:54.02CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures
title DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures
spellingShingle DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures
Stazione, Leonardo
DNA ISOLATION
SCOLYTINAE
DNA QUALITY AND QUANTITY
INSECT SIZE
title_short DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures
title_full DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures
title_fullStr DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures
title_full_unstemmed DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures
title_sort DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures
dc.creator.none.fl_str_mv Stazione, Leonardo
Lantschner, María Victoria
Corley, Juan Carlos
Soliani, Carolina
author Stazione, Leonardo
author_facet Stazione, Leonardo
Lantschner, María Victoria
Corley, Juan Carlos
Soliani, Carolina
author_role author
author2 Lantschner, María Victoria
Corley, Juan Carlos
Soliani, Carolina
author2_role author
author
author
dc.subject.none.fl_str_mv DNA ISOLATION
SCOLYTINAE
DNA QUALITY AND QUANTITY
INSECT SIZE
topic DNA ISOLATION
SCOLYTINAE
DNA QUALITY AND QUANTITY
INSECT SIZE
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.1
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Molecular tools are increasingly used in entomology for several applications such as taxonomy, genetics, ecology and evolution. For these studies, DNA extraction from individual insects is a crucial step, since the yield and quality vary depending on the methods used. Finding an ideal balance between quality and yield is particularly difficult to reach and several constraints must be considered upon selecting the final protocol. Bark beetles (Coleoptera: Curculionidae, Scolytinae) are a diverse group of forest insects for which molecular studies at the individual level are important, given that many species are invasive and may become pests. However, DNA extraction in bark beetles is not easy, given their small size and complex molecules of diverse nature conforming their exoskeleton and present in their digestive tract. Here, we carried out a comparative analysis of DNA extraction performance in five bark beetle species: Hylurgus ligniperda, Hylastes ater, Orthotomicus erosus, Orthotomicus laricis and Cyrtogenius luteus. By assessing the efficiency of two different protocols, our aim was to establish the best species-specific method for population-level studies. Our results showed that a method’s whole performance mainly depends on the species considered and translates into DNA quantity and quality variance. We also noted that the traditional method showed better PCR efficiency for the smallest species whereas the commercial kit performed better for the larger beetles. Our comparative analysis provides evidence that no single method for DNA isolation is best, and that each particular species requires optimization.
Fil: Stazione, Leonardo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina. Università degli Studi di Firenze; Italia
Fil: Lantschner, María Victoria. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina
Fil: Corley, Juan Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina
Fil: Soliani, Carolina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina
description Molecular tools are increasingly used in entomology for several applications such as taxonomy, genetics, ecology and evolution. For these studies, DNA extraction from individual insects is a crucial step, since the yield and quality vary depending on the methods used. Finding an ideal balance between quality and yield is particularly difficult to reach and several constraints must be considered upon selecting the final protocol. Bark beetles (Coleoptera: Curculionidae, Scolytinae) are a diverse group of forest insects for which molecular studies at the individual level are important, given that many species are invasive and may become pests. However, DNA extraction in bark beetles is not easy, given their small size and complex molecules of diverse nature conforming their exoskeleton and present in their digestive tract. Here, we carried out a comparative analysis of DNA extraction performance in five bark beetle species: Hylurgus ligniperda, Hylastes ater, Orthotomicus erosus, Orthotomicus laricis and Cyrtogenius luteus. By assessing the efficiency of two different protocols, our aim was to establish the best species-specific method for population-level studies. Our results showed that a method’s whole performance mainly depends on the species considered and translates into DNA quantity and quality variance. We also noted that the traditional method showed better PCR efficiency for the smallest species whereas the commercial kit performed better for the larger beetles. Our comparative analysis provides evidence that no single method for DNA isolation is best, and that each particular species requires optimization.
publishDate 2024
dc.date.none.fl_str_mv 2024-04
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/265119
Stazione, Leonardo; Lantschner, María Victoria; Corley, Juan Carlos; Soliani, Carolina; DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures; Cold Spring Harbor Laboratory Press; bioRxiv; 4-2024; 1-26
2692-8205
CONICET Digital
CONICET
url http://hdl.handle.net/11336/265119
identifier_str_mv Stazione, Leonardo; Lantschner, María Victoria; Corley, Juan Carlos; Soliani, Carolina; DNA isolation in bark beetles: Reliability of extraction methods and application in downstream molecular procedures; Cold Spring Harbor Laboratory Press; bioRxiv; 4-2024; 1-26
2692-8205
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2024.04.06.588396v1.article-info
info:eu-repo/semantics/altIdentifier/doi/10.1101/2024.04.06.588396
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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