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 | Acceso al texto completo restringido a Biblioteca INIA Las Brujas. Por información adicional contacte bibliolb@inia.org.uy. |
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Registro completo
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Biblioteca (s) : |
INIA Las Brujas. |
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Fecha : |
06/05/2021 |
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Actualizado : |
10/08/2021 |
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Tipo de producción científica : |
Artículos en Revistas Indexadas Internacionales |
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Autor : |
NARANCIO, R.; JOHN, U.; MASON, J.; GIRALDO, P.; SPANGENBERG, G. |
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Afiliación : |
RAFAEL NARANCIO FERES, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; School of Applied Systems Biology, La Trobe University, Melbourne, VIC, Australia; Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia; ULRIK JOHN, Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia; JOHN MASON, School of Applied Systems Biology, La Trobe University, Melbourne, VIC, Australia; Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia; PAULA GIRALDO, Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia; Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Melbourne, VIC, Australia; GERMAN SPANGENBERG, School of Applied Systems Biology, La Trobe University, Melbourne, VIC, Australia; Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia. |
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Título : |
Digital PCR (dPCR) and qPCR mediated determination of transgene copy number in the forage legume white clover (Trifolium repens). |
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Fecha de publicación : |
2021 |
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Fuente / Imprenta : |
Molecular Biology Reports, 2021, volume 48, Issue 4, Pages 3069 - 3077. Doi: https://doi.org/10.1007/s11033-021-06354-5 |
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ISSN : |
0301-4851 |
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DOI : |
10.1007/s11033-021-06354-5 |
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Idioma : |
Inglés |
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Notas : |
Article history: Received, 03 February 2021; Accepted, 12 April 2021; Published, 16 April 2021.
This project was funded by Agriculture Victoria, and Dairy Australia. |
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Contenido : |
ABSTRACT.
Obtaining data on transgene copy number is an integral step in the generation of transgenic plants. Techniques such as Southern blot, segregation analysis, and quantitative PCR (qPCR) have routinely been used for this task, in a range of species. More recently, use of Digital PCR (dPCR) has become prevalent, with a measurement accuracy higher than qPCR reported. Here, the relative merits of qPCR and dPCR for transgene copy number estimation in white clover were investigated. Furthermore, given that single copy reference genes are desirable for estimating gene copy number by relative quantification, and that no single-copy genes have been reported in this species, a search and evaluation of suitable reference genes in white clover was undertaken. Results demonstrated a higher accuracy of dPCR relative to qPCR for copy number estimation in white clover. Two genes, Pyruvate dehydrogenase (PDH), and an ATP-dependent protease, identified as single-copy genes, were used as references for copy number estimation by relative quantification. Identification of single-copy genes in white clover will enable the application of relative quantification for copy number estimation of other genes or transgenes in the species. The results generated here validate the use of dPCR as a reliable strategy for transgene copy number estimation in white clover, and provide resources for future copy number studies in this species.
© 2021, The Author(s), under exclusive licence to Springer Nature B.V. MenosABSTRACT.
Obtaining data on transgene copy number is an integral step in the generation of transgenic plants. Techniques such as Southern blot, segregation analysis, and quantitative PCR (qPCR) have routinely been used for this task, in a range of species. More recently, use of Digital PCR (dPCR) has become prevalent, with a measurement accuracy higher than qPCR reported. Here, the relative merits of qPCR and dPCR for transgene copy number estimation in white clover were investigated. Furthermore, given that single copy reference genes are desirable for estimating gene copy number by relative quantification, and that no single-copy genes have been reported in this species, a search and evaluation of suitable reference genes in white clover was undertaken. Results demonstrated a higher accuracy of dPCR relative to qPCR for copy number estimation in white clover. Two genes, Pyruvate dehydrogenase (PDH), and an ATP-dependent protease, identified as single-copy genes, were used as references for copy number estimation by relative quantification. Identification of single-copy genes in white clover will enable the application of relative quantification for copy number estimation of other genes or transgenes in the species. The results generated here validate the use of dPCR as a reliable strategy for transgene copy number estimation in white clover, and provide resources for future copy number studies in this species.
© 2021, The Author(s), under exclusive licence to Springer Nat... Presentar Todo |
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Palabras claves : |
DdPCR; Reference genes; Relative quantification; Single copy genes; Trifolium repens; Zygosity determination. |
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Asunto categoría : |
F30 Genética vegetal y fitomejoramiento |
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Marc : |
LEADER 02569naa a2200277 a 4500
001 1062022
005 2021-08-10
008 2021 bl uuuu u00u1 u #d
022 $a0301-4851
024 7 $a10.1007/s11033-021-06354-5$2DOI
100 1 $aNARANCIO, R.
245 $aDigital PCR (dPCR) and qPCR mediated determination of transgene copy number in the forage legume white clover (Trifolium repens).$h[electronic resource]
260 $c2021
500 $aArticle history: Received, 03 February 2021; Accepted, 12 April 2021; Published, 16 April 2021. This project was funded by Agriculture Victoria, and Dairy Australia.
520 $aABSTRACT. Obtaining data on transgene copy number is an integral step in the generation of transgenic plants. Techniques such as Southern blot, segregation analysis, and quantitative PCR (qPCR) have routinely been used for this task, in a range of species. More recently, use of Digital PCR (dPCR) has become prevalent, with a measurement accuracy higher than qPCR reported. Here, the relative merits of qPCR and dPCR for transgene copy number estimation in white clover were investigated. Furthermore, given that single copy reference genes are desirable for estimating gene copy number by relative quantification, and that no single-copy genes have been reported in this species, a search and evaluation of suitable reference genes in white clover was undertaken. Results demonstrated a higher accuracy of dPCR relative to qPCR for copy number estimation in white clover. Two genes, Pyruvate dehydrogenase (PDH), and an ATP-dependent protease, identified as single-copy genes, were used as references for copy number estimation by relative quantification. Identification of single-copy genes in white clover will enable the application of relative quantification for copy number estimation of other genes or transgenes in the species. The results generated here validate the use of dPCR as a reliable strategy for transgene copy number estimation in white clover, and provide resources for future copy number studies in this species. © 2021, The Author(s), under exclusive licence to Springer Nature B.V.
653 $aDdPCR
653 $aReference genes
653 $aRelative quantification
653 $aSingle copy genes
653 $aTrifolium repens
653 $aZygosity determination
700 1 $aJOHN, U.
700 1 $aMASON, J.
700 1 $aGIRALDO, P.
700 1 $aSPANGENBERG, G.
773 $tMolecular Biology Reports, 2021, volume 48, Issue 4, Pages 3069 - 3077. Doi: https://doi.org/10.1007/s11033-021-06354-5
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INIA Las Brujas (LB) |
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| 11. |  | PEREZ GOMAR, E.; BEMHAJA, M.; GUTIÉRREZ, F. Estudio exploratorio sobre las propiedades químicas de suelos de sistemas silvopastoriles de la Unidad Chapicuy en Paysandú, Uruguay. ln: CONGRESO NACIONAL DE SISTEMAS SILVOPASTORILES, 1., 2009, Posadas, Misiones, AR. [Posters]. Buenos Aires: INTA, 2009.| Biblioteca(s): INIA La Estanzuela. |
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| 12. | | Pérez Gomar, E.; Bemhaja, M.; Gutiérrez, F. Evaluación de la vegetación, biomasa radicular y propiedades físico químcas del suelo en sistemas pastoriles y forestales en suelos arenosos de Tacuarembó ln: Reunión del grupo técnico en forrajeras del Cono Sur, 22., 2008, Minas, Uruguay Bioma campos: innovando para mantener su sustentabilidad y competitividad. Memorias. Minas (Uruguay): INIA; FAO; PROCISUR, 2008. p. 186 Versión impresa y en CD ROM Instituto Nacional de Investigación Agropecuaria, Uruguay; FAO; PROCISUR| Biblioteca(s): INIA Las Brujas; INIA Tacuarembó. |
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| 20. | | PEREZ GOMAR, E.; BEMHAJA SARAIVA, M.; GUTIERREZ, F. Estudio exploratorio sobre las propiedades químicas de suelos de sistemas silvopastoriles de la Unidad Chapicuy en Paysandú, Uruguay. ln: Congreso Nacional de Sistemas Silvopastoriles, 1., 2009, Posadas, Misiones, Argentina Actas. Buenos Aires (Argentina): INTA, 2009. p. 450-452| Tipo: Trabajos en Congresos/Conferencias |
| Biblioteca(s): INIA Tacuarembó. |
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| Registros recuperados : 38 | |
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