Direct PCR using Spodoptera frugiperda eggs as template

Autores
Loto, Flavia del Valle; Baigori, Mario Domingo; Pera, Licia Maria
Año de publicación
2008
Idioma
español castellano
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Introduction: PCR is widely employed in a variety of experimental applications such as studies of Spodoptera frugiperda (Sf), an important pest in America. Objective: To optimize direct PCR technique using Sf samples. Material and methods: PCR reactions were performed in 25 μL containing: 21.4 μl sterile water, 0.1 μl of each oligonucleotide primers (JM76 and JM77), 5 μl 5X Taq buffer and 0.2 μl Taq DNA polymerase. One, two or three eggs were added to the reaction mixture. Negative and positive controls were performed with distilled water and purified DNA, respectively. Amplification was performed with the following program: An initial denaturation step at 97ºC for 5 min and then 35 cycles of the following: 1 min at 94ºC, 1 min at 58ºC, 2 min at 72ºC, and a final extension at 72ºC 2 min. Results and Discussion: Samples with 2 eggs were amplified with the same quality as samples amplified with purified DNA. Only one of three amplifications assays was positive in samples with one egg. Samples with 3 eggs were not amplified, probably due to excessive DNA or PCR inhibitors. The method is simple, fast and cost saving. This work was supported by grants PICTO-UNT 761 and PIP 6062 (CONICET).
Fil: Loto, Flavia del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
Fil: Baigori, Mario Domingo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
Fil: Pera, Licia Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
XXV Annual Scientific Meeting Tucuman Biology Association
Tafi del Valle
Argentina
Asociación de Biología de Tucumán
Materia
PCR DIRECTA
SPODOPTERA FRUGIPERDA
HUEVOS
BIOTIPOS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/199466
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/199466
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Direct PCR using Spodoptera frugiperda eggs as templateLoto, Flavia del ValleBaigori, Mario DomingoPera, Licia MariaPCR DIRECTASPODOPTERA FRUGIPERDAHUEVOSBIOTIPOShttps://purl.org/becyt/ford/2.11https://purl.org/becyt/ford/2Introduction: PCR is widely employed in a variety of experimental applications such as studies of Spodoptera frugiperda (Sf), an important pest in America. Objective: To optimize direct PCR technique using Sf samples. Material and methods: PCR reactions were performed in 25 μL containing: 21.4 μl sterile water, 0.1 μl of each oligonucleotide primers (JM76 and JM77), 5 μl 5X Taq buffer and 0.2 μl Taq DNA polymerase. One, two or three eggs were added to the reaction mixture. Negative and positive controls were performed with distilled water and purified DNA, respectively. Amplification was performed with the following program: An initial denaturation step at 97ºC for 5 min and then 35 cycles of the following: 1 min at 94ºC, 1 min at 58ºC, 2 min at 72ºC, and a final extension at 72ºC 2 min. Results and Discussion: Samples with 2 eggs were amplified with the same quality as samples amplified with purified DNA. Only one of three amplifications assays was positive in samples with one egg. Samples with 3 eggs were not amplified, probably due to excessive DNA or PCR inhibitors. The method is simple, fast and cost saving. This work was supported by grants PICTO-UNT 761 and PIP 6062 (CONICET).Fil: Loto, Flavia del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Baigori, Mario Domingo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Pera, Licia Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaXXV Annual Scientific Meeting Tucuman Biology AssociationTafi del ValleArgentinaAsociación de Biología de TucumánTech Science Press2008info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/199466Direct PCR using Spodoptera frugiperda eggs as template; XXV Annual Scientific Meeting Tucuman Biology Association; Tafi del Valle; Argentina; 2008; 152-1520327-9545CONICET DigitalCONICETspainfo:eu-repo/semantics/altIdentifier/url/http://www.asobioltuc.com/uploads/archivos/1503954099_QmlvY2VsbCBYWFZJIEpDIDIwMDkucGRm.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:04:38Zoai:ri.conicet.gov.ar:11336/199466instacron: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:34982025-09-03 10:04:39.069CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Direct PCR using Spodoptera frugiperda eggs as template
title Direct PCR using Spodoptera frugiperda eggs as template
spellingShingle Direct PCR using Spodoptera frugiperda eggs as template
Loto, Flavia del Valle
PCR DIRECTA
SPODOPTERA FRUGIPERDA
HUEVOS
BIOTIPOS
title_short Direct PCR using Spodoptera frugiperda eggs as template
title_full Direct PCR using Spodoptera frugiperda eggs as template
title_fullStr Direct PCR using Spodoptera frugiperda eggs as template
title_full_unstemmed Direct PCR using Spodoptera frugiperda eggs as template
title_sort Direct PCR using Spodoptera frugiperda eggs as template
dc.creator.none.fl_str_mv Loto, Flavia del Valle
Baigori, Mario Domingo
Pera, Licia Maria
author Loto, Flavia del Valle
author_facet Loto, Flavia del Valle
Baigori, Mario Domingo
Pera, Licia Maria
author_role author
author2 Baigori, Mario Domingo
Pera, Licia Maria
author2_role author
author
dc.subject.none.fl_str_mv PCR DIRECTA
SPODOPTERA FRUGIPERDA
HUEVOS
BIOTIPOS
topic PCR DIRECTA
SPODOPTERA FRUGIPERDA
HUEVOS
BIOTIPOS
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.11
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv Introduction: PCR is widely employed in a variety of experimental applications such as studies of Spodoptera frugiperda (Sf), an important pest in America. Objective: To optimize direct PCR technique using Sf samples. Material and methods: PCR reactions were performed in 25 μL containing: 21.4 μl sterile water, 0.1 μl of each oligonucleotide primers (JM76 and JM77), 5 μl 5X Taq buffer and 0.2 μl Taq DNA polymerase. One, two or three eggs were added to the reaction mixture. Negative and positive controls were performed with distilled water and purified DNA, respectively. Amplification was performed with the following program: An initial denaturation step at 97ºC for 5 min and then 35 cycles of the following: 1 min at 94ºC, 1 min at 58ºC, 2 min at 72ºC, and a final extension at 72ºC 2 min. Results and Discussion: Samples with 2 eggs were amplified with the same quality as samples amplified with purified DNA. Only one of three amplifications assays was positive in samples with one egg. Samples with 3 eggs were not amplified, probably due to excessive DNA or PCR inhibitors. The method is simple, fast and cost saving. This work was supported by grants PICTO-UNT 761 and PIP 6062 (CONICET).
Fil: Loto, Flavia del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
Fil: Baigori, Mario Domingo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
Fil: Pera, Licia Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
XXV Annual Scientific Meeting Tucuman Biology Association
Tafi del Valle
Argentina
Asociación de Biología de Tucumán
description Introduction: PCR is widely employed in a variety of experimental applications such as studies of Spodoptera frugiperda (Sf), an important pest in America. Objective: To optimize direct PCR technique using Sf samples. Material and methods: PCR reactions were performed in 25 μL containing: 21.4 μl sterile water, 0.1 μl of each oligonucleotide primers (JM76 and JM77), 5 μl 5X Taq buffer and 0.2 μl Taq DNA polymerase. One, two or three eggs were added to the reaction mixture. Negative and positive controls were performed with distilled water and purified DNA, respectively. Amplification was performed with the following program: An initial denaturation step at 97ºC for 5 min and then 35 cycles of the following: 1 min at 94ºC, 1 min at 58ºC, 2 min at 72ºC, and a final extension at 72ºC 2 min. Results and Discussion: Samples with 2 eggs were amplified with the same quality as samples amplified with purified DNA. Only one of three amplifications assays was positive in samples with one egg. Samples with 3 eggs were not amplified, probably due to excessive DNA or PCR inhibitors. The method is simple, fast and cost saving. This work was supported by grants PICTO-UNT 761 and PIP 6062 (CONICET).
publishDate 2008
dc.date.none.fl_str_mv 2008
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/199466
Direct PCR using Spodoptera frugiperda eggs as template; XXV Annual Scientific Meeting Tucuman Biology Association; Tafi del Valle; Argentina; 2008; 152-152
0327-9545
CONICET Digital
CONICET
url http://hdl.handle.net/11336/199466
identifier_str_mv Direct PCR using Spodoptera frugiperda eggs as template; XXV Annual Scientific Meeting Tucuman Biology Association; Tafi del Valle; Argentina; 2008; 152-152
0327-9545
CONICET Digital
CONICET
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language spa
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dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Tech Science Press
publisher.none.fl_str_mv Tech Science 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 13.13397

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