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| 3. |  | ALTIER, N. Enfermedades de alfalfa y estrategias para su manejo In: INIA (Instituto Nacional de Investigación Agropecuaria); MGAP (Ministerio Ganadería, Agricultura y Pesca); IPA (Instituto Plan Agropecuario). Jornada de Alfalfa: Todo lo que usted quiería saber sobre la reina de las forrajeras. (Jornada de Alfalfa, Colonia, Uruguay, 1998). Colonia (Uruguay): INIA, MGAP, Plan Agropecuario, 1998. p. 9-14 INIA; MGAP| Biblioteca(s): INIA Las Brujas. |
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| 7. | | ALTIER, N. Enfermedades de pasturas. ln: Altier, N.; Rebuffo, M.; Cabrera, K. (eds.). Enfermedades y plagas en pasturas. Montevideo (Uruguay): INIA, 2010. p.19-35 (INIA Serie Técnica; 183) Contiene diagrama de las enfermedades en pasturas| Biblioteca(s): INIA Las Brujas; INIA Tacuarembó. |
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| 10. | | ALTIER, N. Impacto de las enfermedades en la producción de pasturas In: RISSO, D.; BERRETTA, E.J.; MORON, A. (Eds.). Producción y manejo de pasturas. Seminario técnico, 17-19 octubre 1995, INIA Tacuarembó. Montevideo (Uruguay): INIA, 1996. p47-56 (INIA Serie Técnica; 80)| Biblioteca(s): INIA La Estanzuela; INIA Las Brujas; INIA Tacuarembó. |
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| 14. | | GARAYCOCHEA, S.; ALTIER, N. Estudio de la diversidad genética de las comunidades microbianas asociadas a la dinámica del P en suelos de Uruguay. GGM 47 - COMUNICACIONES LIBRES - GGM. GENÓMICA Y GENÉTICA MOLECULAR In: JOURNAL OF BASIC & APPLIED GENETICS, 2016, Vol.27, Iss. 1 (Supp.). XVI LATIN AMERICAN CONGRESS OF GENETICS, IV CONGRESS OF THE URUGUAYAN SOCIETY OF GENETICS, XLIX ANNUAL MEETING OF THE GENETICS SOCIETY OF CHILE, XLV ARGENTINE CONGRESS OF GENETICS, 9-12 October 2016. PROCEEDINGS. Montevideo (Uruguay): SAG, 2016. p. 278| Biblioteca(s): INIA Las Brujas. |
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| 18. | | REBUFFO, M.; ALTIER, N. Mejoramiento genetico de trébol rojo In: RISSO, D.; BERRETTA, E.J.; MORON, A. (Eds.). Producción y manejo de pasturas. Seminario técnico, 17-19 octubre 1995, INIA Tacuarembó. Montevideo (Uruguay): INIA, 1996. p151-153 (INIA Serie Técnica; 80)| Biblioteca(s): INIA La Estanzuela; INIA Las Brujas; INIA Tacuarembó. |
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Registro completo
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Biblioteca (s) : |
INIA Las Brujas. |
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Fecha actual : |
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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Circulación / Nivel : |
Internacional - -- |
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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
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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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