Principales herramientas moleculares empleadas en la ciencia animal
- Autores:
-
ARANGUREN, YANI
MACHADO, ELWI
MONTES V., DONICER
- Tipo de recurso:
- Article of journal
- Fecha de publicación:
- 2013
- Institución:
- Universidad de Sucre
- Repositorio:
- Repositorio Unisucre
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unisucre.edu.co:001/1452
- Acceso en línea:
- https://repositorio.unisucre.edu.co/handle/001/1452
https://doi.org/10.24188/recia.v5.n2.2013.459
- Palabra clave:
- diagnostics
DNA
Genetic Variation
PCR.
diagnostico
DNA
PCR
variación genética.
- Rights
- openAccess
- License
- https://creativecommons.org/licenses/by-nc-sa/4.0/
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dc.title.spa.fl_str_mv |
Principales herramientas moleculares empleadas en la ciencia animal |
dc.title.translated.eng.fl_str_mv |
Main molecular tools employed in animal science |
title |
Principales herramientas moleculares empleadas en la ciencia animal |
spellingShingle |
Principales herramientas moleculares empleadas en la ciencia animal diagnostics DNA Genetic Variation PCR. diagnostico DNA PCR variación genética. |
title_short |
Principales herramientas moleculares empleadas en la ciencia animal |
title_full |
Principales herramientas moleculares empleadas en la ciencia animal |
title_fullStr |
Principales herramientas moleculares empleadas en la ciencia animal |
title_full_unstemmed |
Principales herramientas moleculares empleadas en la ciencia animal |
title_sort |
Principales herramientas moleculares empleadas en la ciencia animal |
dc.creator.fl_str_mv |
ARANGUREN, YANI MACHADO, ELWI MONTES V., DONICER |
dc.contributor.author.spa.fl_str_mv |
ARANGUREN, YANI MACHADO, ELWI MONTES V., DONICER |
dc.subject.eng.fl_str_mv |
diagnostics DNA Genetic Variation PCR. |
topic |
diagnostics DNA Genetic Variation PCR. diagnostico DNA PCR variación genética. |
dc.subject.spa.fl_str_mv |
diagnostico DNA PCR variación genética. |
publishDate |
2013 |
dc.date.accessioned.none.fl_str_mv |
2013-07-12 00:00:00 2022-07-01T17:15:36Z |
dc.date.available.none.fl_str_mv |
2013-07-12 00:00:00 2022-07-01T17:15:36Z |
dc.date.issued.none.fl_str_mv |
2013-07-12 |
dc.type.spa.fl_str_mv |
Artículo de revista |
dc.type.eng.fl_str_mv |
Journal article |
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http://purl.org/coar/resource_type/c_2df8fbb1 |
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10.24188/recia.v5.n2.2013.459 |
dc.identifier.eissn.none.fl_str_mv |
2027-4297 |
dc.identifier.url.none.fl_str_mv |
https://doi.org/10.24188/recia.v5.n2.2013.459 |
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https://repositorio.unisucre.edu.co/handle/001/1452 https://doi.org/10.24188/recia.v5.n2.2013.459 |
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10.24188/recia.v5.n2.2013.459 2027-4297 |
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ANTONANGELO, A.; COLOMBI, D.; CURI, R.; BRAZ, A.; OLIVEIRA, T. y MOTAL, M. 2012. Detection and quantification of Duffy antigen on bovine red blood cell membranes using a polyclonal antibody. Pesq. Vet. Bras. 32(9): 936–940. APARICIO, S.; CHAPMAN, J.; STUPKA, E.; PUTNAM, N.; CHIA, J.; DEHAL, P.; CHRISTOFFELS, A. et al . Whole-genome shotgun assembly and analysis of the genome of Fugu rubripes. Science 297(5585): 1301–1310. AZOFEIFA, A. 2006. Uso de marcadores moleculares en plantas; aplicaciones en frutales del trópico. Agronomía Mesoamericana, 17: 221-242. BAUER, O. 1976. Fitogenética aplicada. Editorial Limusa. México D.F. BENSON, D.; KARSCH-MIZRACHI, I.; LIPMAN, D.; OSTELL, J. y WHEELER, D. 2005. GenBank. Nucleic acids research 33: D34–D38. BOTSTEIN, D.; WHITE, R.; SKOLNICK, M. y DAVIS, R. 1980. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. The American Journal of Human Genetics 32: 314-331. BOVINE GENOME SEQUENCING AND ANALYSIS CONSORTIUM. 2009. The genome sequence of taurine cattle: a window to ruminant biology and evolution. Science 324(5926):522-528. BOVINE HAPMAP CONSORTIUM. 2009. Genome-wide survey of SNP variation uncovers the genetic structure of cattle breeds. Science 324(5926):528-532. BREYNE, P.; DREESEN, R.; CANNOOT, B.; ROMBAUT, D.; VANDEPOELE, K.; ROMBAUTS, S.; VANDERHAEGHEN, R.; INZÉ, D. y ZABEAU, M. 2003. Quantitative cDNA-AFLP analysis for genome-wide expression studies. Molecular genetics and genomics 269: 173-179. CARLEER, J.; PASTORET, P. y ANSAY, M. 1978. Isozyme characterization of cattle (Bos taurus) and American buffalo (Bison bison) cell cultures. Animal Blood Groups Biochemical Genetics 9(3):175-179. DA SILVA, J.; ANDRÉ, M.; DA FONSECA, A.; LOPES, C.; DA SILVA, D.; DE ANDRADE, S.; OLIVEIRA, C. y BARBOSA, J. 2013. Molecular and serological prevalence of Babesia bovis and Babesia bigemina in water buffaloes in the north region of Brazil. Veterinary parasitology 197(3-4): 678-681. DEKKERS, J.C.M. 2004. Commercial application of marker and gene assisted selection in livestock: Strategies and lessons. Journal of Animal Science, v.82, p.313-328 DALE, J. y VON SCHANTZ, M. 2002. From Genes to Genomes: Concepts and Applications of DNA Technology. Editorial John Wiley y Sons, Ltd. Londres. DJARI, A.; ESQUERRÉ, D., WEISS, B., MARTINS, F., MEERSSEMAN, C., BOUSSAHA, M., KLOPP, C. y ROCHA D. 2013.Gene-based single nucleotide polymorphism discovery in bovine muscle using next-generation transcriptomic sequencing. BMC Genomics 14:307. DI BELLA, J.; BAO, Y.; GLOOR, G.; BURTON, J. y REID, G. 2013. High throughput sequencing methods and analysis for microbiome research. Journal of microbiological methods 5(3): 401-414. DUAN, Z.; GUO, Y. y LIU, J. 2006. Application of modern molecular biology techniques to study micro-ecosystem in the rumen. Wei Sheng Wu Xue Bao 46: 166-169. FERREIRA, E.; DE LANA, M.; CARNEIRO, M.; REIS, A.; PAES, D.; DA SILVA, E.; SCHALLIG, H. y GONTIJO, C. 2007. Comparison of serological assays for the diagnosis of canine visceral leishmaniasis in animals presenting different clinical manifestations. Veterinary parasitology 146(3-4): 235-241. FRANK, D. y PACE, N. 2008. Gastrointestinal microbiology enters the metagenomics era. Current opinion in gastroenterology 24(1): 4-10. FRANKHAM, R.; BALLOU, J. y BRISCOE, D. 2004. Introduction to conservation genetics. Editorial Cambridge University Press, Cambridge. GEORGES, M. 2012. Impact of high-throughput genotyping and sequencing on the identification of genes and variants underlying phenotypic variation in domestic cattle. En: Womack, J. (eds). Bovine Genomic. Editorial John Wiley y Sons, Ltd, Oxford. HEBERT, P.; CYWINSKA, A.; BALL, S. y DE WAARD, J. 2003. Biological identifications through DNA barcodes. Proceedings of the Royal Society B 270: 313-321. HILLIER, W.; MILLER, W.; BIRNEY, E.; WARREN, W.; HARDISON, R.; PONTING C.; et al . 2004. Sequence and comparative analysis of the chicken genome provide unique perspectives on vertebrate evolution. Nature 432(7018): 695-716. HOBSON, P. y STEWART, C. (eds). 1997. The Rumen Microbial Ecosystem (2da edicion). Editorial Springer Netherlands, Dordrecht, Holanda. HUNGATE, R. 1966. The rumen and its microbes. Editorial Academic Press, New York. HYTEN, D.; SONG, Q.; FICKUS, E.; QUIGLEY, C.; LIM, J.; CHOI, I.; HWANG, E.; PASTOR-CORRALES, M. y CREGAN, P. 2010. High-throughput SNP discovery and assay development in common bean. BMC genomics 11: 475-483. INTERNATIONAL CHICKEN GENOME SEQUENCING CONSORTIUM. 2004. Sequence and comparative analysis of the chicken genome provide unique perspectives on vertebrate evolution. Nature 432: 695-716. KIM, M.; MORRISON, M. y YU, Z. 2011. Status of the phylogenetic diversity census of ruminal microbiomes. FEMS microbiology ecology 76(1): 49-63. LIPMAN,D. National Center for Biotechnology information. [serial online] 2013 [Citado 30 Dic 2013]. Disponible en: http://www.ncbi.nlm.nih.gov/snp/?term=Bos taurus. LITT, M. y LUTY, J. 1989. A hypervariable microsatellite revealed by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene. The American Journal of Human Genetics 44:397-401. LUQUE, J. y HERRÁEZ, A. 2001. Texto ilustrado de biología molecular e ingeniería genética: conceptos, técnicas y aplicaciones en ciencias de la salud. Editorial Elsevier. Madrid. MASKOS, U. y SOUTHERN, E. 1992. Oligonucleotide hybridisations on glass supports: a novel linker for oligonucleotide synthesis and hybridisation properties of oligonucleotides synthesised in situ. Nucleic Acids Research 20(7): 1679-1684. MATSUZAKI, H.; LOI, H.; DONG, S.; TSAI, Y.; FANG, J.; LAW, J.; DI, X.; et al., 2004. Parallel genotyping of over 10,000 SNPs using a one-primer assay on a high-density oligonucleotide array. Genome research 14(3): 414-25. MATUKUMALLI, L.; LAWLEY, C.; SCHNABEL, R.; TAYLOR, J.; ALLAN, M.; HEATON, M.; O’CONNELL, J.; MOORE, S.; SMITH, T.; SONSTEGARD, T. y VAN TASSELL, C. 2009. Development and characterization of a high density SNP genotyping assay for cattle. PloS one 4(4): e5350 Mac SWEENEY, C. y MACKIE, R. 2012. Micro-organisms and ruminant digestion: state of knowledge, trands and future prospects. Commission On Genetic Resources for Food and Agriculture (Editores), Roma: disponible en: http://www.fao.org/docrep/016/ap099e/ap099e.pdf. METZKER, M. 2010. Sequencing technologies - the next generation. Nature reviews. Genetics 11(1): 31-46. MONIS, P.; GIGLIO, S.; KEEGAN, A. y THOMPSON, R. 2005. Emerging technologies for the detection and genetic characterization of protozoan parasites. Trends Parasitol. 21:340-346. MULLIS, K.; FALOONA, F.; SCHARF, S.; SAIKI, R.; HORN, G. y ERLICH, H. 1986. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Old Spring Harbor Symposia on Quantitative Biology 51(1):263-73. NELSON, M,; MARNELLOS, G.; KAMMERER, S.; HOYAL, C.; SHI, M.; CANTOR, C. y BRAUN, A. 2004. Large-Scale Validation of Single Nucleotide Polymorphisms in Gene Regions. Genome Research 14(8): 1664-1668. OSOEGAWA, K.; WOON, P.; ZHAO, B.; FRENGEN, E.; TATENO, M.; CATANESE, J. y DE JONG, P. 1998. An improved approach for construction of bacterial artificial chromosome libraries. Genomics 52(1): 1-8. PAREEK, C.; SMOCZYNSKI, R. y TRETYN, A. 2011. Sequencing technologies and genome sequencing. Journal of applied genetics 52(4): 413-435. POLIDO, P.; FERREIRA, F.; ALBERTON, O. y HÜLSE, S. 2012. Marcadores moleculares aplicados no melhoramento genético de bovinos. Arquivos de Ciências Veterinárias e Zoologia da UNIPAR 15(2): 161-169. SAIKI, R.; GELFAND, D.; STOFFEL, S.; SCHARF, S.; HIGUCHI, R.; HORN, G.; MULLIS, K. y ERLICH, H. 1988. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239(4839): 487-491. SAMBROOK, J., RUSSELL, D. 2001. Molecular Cloning: A Laboratory Manual. (3ra edición) Editorial Cold Spring Harbor Laboratory Press, Nueva York. SANGER, F.; AIR, G.; BARRELL, B.; BROWN, N.; COULSON, A.; FIDDES, J.; HUTCHISON, C.; SLOCOMBE, P. y SMITH, M. 1977. Nucleotide sequence of bacteriophage ?X174 DNA. Nature 265(5596): 687-695. SCHOOK, L.; BEEVER, J.; ROGERS, J.; HUMPHRAY, S.; ARCHIBALD, A.; CHARDON, P.; MILAN, D.; ROHRER, G. y EVERSOLE, K. 2005. Swine Genome Sequencing Consortium (SGSC): a strategic roadmap for sequencing the pig genome. Comparative and Functinal Genomics (4): 251-255. TAUTZ, D. 1989. Hypervariability of simple sequences as a general source for polymorphic DNA markers. Nucleic Acids Research 17:6463-6471. THORNTON, P. 2010. Livestock production: recent trends,future prospects. Philosophical Transactions B 365: 2853-2867. VENTER, J.; ADAMS, D.; MYERS, E.; et al., 2001. The sequence of the human genome. Science 291(5507): 1304-1351. WILLIAMS.; J.; KUBEILIK.; A.; LIVAK.; K.; RAFALSKI.; J. y TINGEY.; S. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research 18:6531-6535. XIAO.; L. 2010. Molecular epidemiology of cryptosporidiosis: an update. Experimental parasitology 124(1): 80-9. YANG.; W.; KANG.; X. ; YANG.; Q.; LIN.; Y. y FANG.; M. 2013. Review on the development of genotyping methods for assessing farm animal diversity. Journal of Animal Science and Biotechnology 4: 2-8. ZABEAU.; M. y VOS.; P. 1993. Selective restriction fragment amplification: a general method for DNA fingerprinting. EP Patent. publication. No. 0534858:B2. ZIMIN.; A.; DELCHER.; A.; FLOREA.; L.; KELLEY.; D.; SCHATZ.; M.; PUIU.; D.; HANRAHAN.; F.; PERTEA.; G.; VAN TASSELL.; C.; SONSTEGARD.; T.; MARÇAIS.; G.; ROBERTS.; M.; SUBRAMANIAN.; P.; YORKE.; J. y SALZBERG.; S. 2009. A whole-genome assembly of the domestic cow.; Bos taurus. Genome biology 10(4): R42. |
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ARANGUREN, YANI6c98db485db7f6d3897a920f46a56dec500MACHADO, ELWIbab8651626ec6c0f936b3ca5fb5d84b8500MONTES V., DONICERa66edd7487d2e315d53ce65369f99aed2013-07-12 00:00:002022-07-01T17:15:36Z2013-07-12 00:00:002022-07-01T17:15:36Z2013-07-12https://repositorio.unisucre.edu.co/handle/001/145210.24188/recia.v5.n2.2013.4592027-4297https://doi.org/10.24188/recia.v5.n2.2013.459application/pdfspaUniversidad de Sucrehttps://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2https://revistas.unisucre.edu.co/index.php/recia/article/view/459diagnosticsDNAGenetic VariationPCR.diagnosticoDNAPCRvariación genética.Principales herramientas moleculares empleadas en la ciencia animalMain molecular tools employed in animal scienceArtículo de revistaJournal articleinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/resource_type/c_dcae04bchttp://purl.org/coar/resource_type/c_2df8fbb1Texthttp://purl.org/redcol/resource_type/ARTREVhttp://purl.org/coar/version/c_970fb48d4fbd8a85ANTONANGELO, A.; COLOMBI, D.; CURI, R.; BRAZ, A.; OLIVEIRA, T. y MOTAL, M. 2012. Detection and quantification of Duffy antigen on bovine red blood cell membranes using a polyclonal antibody. Pesq. Vet. Bras. 32(9): 936–940.APARICIO, S.; CHAPMAN, J.; STUPKA, E.; PUTNAM, N.; CHIA, J.; DEHAL, P.; CHRISTOFFELS, A. et al . Whole-genome shotgun assembly and analysis of the genome of Fugu rubripes. Science 297(5585): 1301–1310.AZOFEIFA, A. 2006. Uso de marcadores moleculares en plantas; aplicaciones en frutales del trópico. Agronomía Mesoamericana, 17: 221-242.BAUER, O. 1976. Fitogenética aplicada. Editorial Limusa. México D.F. BENSON, D.; KARSCH-MIZRACHI, I.; LIPMAN, D.; OSTELL, J. y WHEELER, D. 2005. GenBank. Nucleic acids research 33: D34–D38.BOTSTEIN, D.; WHITE, R.; SKOLNICK, M. y DAVIS, R. 1980. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. The American Journal of Human Genetics 32: 314-331.BOVINE GENOME SEQUENCING AND ANALYSIS CONSORTIUM. 2009. The genome sequence of taurine cattle: a window to ruminant biology and evolution. Science 324(5926):522-528.BOVINE HAPMAP CONSORTIUM. 2009. Genome-wide survey of SNP variation uncovers the genetic structure of cattle breeds. Science 324(5926):528-532.BREYNE, P.; DREESEN, R.; CANNOOT, B.; ROMBAUT, D.; VANDEPOELE, K.; ROMBAUTS, S.; VANDERHAEGHEN, R.; INZÉ, D. y ZABEAU, M. 2003. Quantitative cDNA-AFLP analysis for genome-wide expression studies. Molecular genetics and genomics 269: 173-179.CARLEER, J.; PASTORET, P. y ANSAY, M. 1978. Isozyme characterization of cattle (Bos taurus) and American buffalo (Bison bison) cell cultures. Animal Blood Groups Biochemical Genetics 9(3):175-179.DA SILVA, J.; ANDRÉ, M.; DA FONSECA, A.; LOPES, C.; DA SILVA, D.; DE ANDRADE, S.; OLIVEIRA, C. y BARBOSA, J. 2013. Molecular and serological prevalence of Babesia bovis and Babesia bigemina in water buffaloes in the north region of Brazil. Veterinary parasitology 197(3-4): 678-681. DEKKERS, J.C.M. 2004. Commercial application of marker and gene assisted selection in livestock: Strategies and lessons. Journal of Animal Science, v.82, p.313-328DALE, J. y VON SCHANTZ, M. 2002. From Genes to Genomes: Concepts and Applications of DNA Technology. Editorial John Wiley y Sons, Ltd. Londres. DJARI, A.; ESQUERRÉ, D., WEISS, B., MARTINS, F., MEERSSEMAN, C., BOUSSAHA, M., KLOPP, C. y ROCHA D. 2013.Gene-based single nucleotide polymorphism discovery in bovine muscle using next-generation transcriptomic sequencing. BMC Genomics 14:307.DI BELLA, J.; BAO, Y.; GLOOR, G.; BURTON, J. y REID, G. 2013. High throughput sequencing methods and analysis for microbiome research. Journal of microbiological methods 5(3): 401-414.DUAN, Z.; GUO, Y. y LIU, J. 2006. Application of modern molecular biology techniques to study micro-ecosystem in the rumen. Wei Sheng Wu Xue Bao 46: 166-169.FERREIRA, E.; DE LANA, M.; CARNEIRO, M.; REIS, A.; PAES, D.; DA SILVA, E.; SCHALLIG, H. y GONTIJO, C. 2007. Comparison of serological assays for the diagnosis of canine visceral leishmaniasis in animals presenting different clinical manifestations. Veterinary parasitology 146(3-4): 235-241.FRANK, D. y PACE, N. 2008. Gastrointestinal microbiology enters the metagenomics era. Current opinion in gastroenterology 24(1): 4-10.FRANKHAM, R.; BALLOU, J. y BRISCOE, D. 2004. Introduction to conservation genetics. Editorial Cambridge University Press, Cambridge. GEORGES, M. 2012. Impact of high-throughput genotyping and sequencing on the identification of genes and variants underlying phenotypic variation in domestic cattle. En: Womack, J. (eds). Bovine Genomic. Editorial John Wiley y Sons, Ltd, Oxford.HEBERT, P.; CYWINSKA, A.; BALL, S. y DE WAARD, J. 2003. Biological identifications through DNA barcodes. Proceedings of the Royal Society B 270: 313-321.HILLIER, W.; MILLER, W.; BIRNEY, E.; WARREN, W.; HARDISON, R.; PONTING C.; et al . 2004. Sequence and comparative analysis of the chicken genome provide unique perspectives on vertebrate evolution. Nature 432(7018): 695-716.HOBSON, P. y STEWART, C. (eds). 1997. The Rumen Microbial Ecosystem (2da edicion). Editorial Springer Netherlands, Dordrecht, Holanda.HUNGATE, R. 1966. The rumen and its microbes. Editorial Academic Press, New York.HYTEN, D.; SONG, Q.; FICKUS, E.; QUIGLEY, C.; LIM, J.; CHOI, I.; HWANG, E.; PASTOR-CORRALES, M. y CREGAN, P. 2010. High-throughput SNP discovery and assay development in common bean. BMC genomics 11: 475-483.INTERNATIONAL CHICKEN GENOME SEQUENCING CONSORTIUM. 2004. Sequence and comparative analysis of the chicken genome provide unique perspectives on vertebrate evolution. Nature 432: 695-716.KIM, M.; MORRISON, M. y YU, Z. 2011. Status of the phylogenetic diversity census of ruminal microbiomes. FEMS microbiology ecology 76(1): 49-63.LIPMAN,D. National Center for Biotechnology information. [serial online] 2013 [Citado 30 Dic 2013]. Disponible en: http://www.ncbi.nlm.nih.gov/snp/?term=Bos taurus.LITT, M. y LUTY, J. 1989. A hypervariable microsatellite revealed by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene. The American Journal of Human Genetics 44:397-401.LUQUE, J. y HERRÁEZ, A. 2001. Texto ilustrado de biología molecular e ingeniería genética: conceptos, técnicas y aplicaciones en ciencias de la salud. Editorial Elsevier. Madrid.MASKOS, U. y SOUTHERN, E. 1992. Oligonucleotide hybridisations on glass supports: a novel linker for oligonucleotide synthesis and hybridisation properties of oligonucleotides synthesised in situ. Nucleic Acids Research 20(7): 1679-1684.MATSUZAKI, H.; LOI, H.; DONG, S.; TSAI, Y.; FANG, J.; LAW, J.; DI, X.; et al., 2004. Parallel genotyping of over 10,000 SNPs using a one-primer assay on a high-density oligonucleotide array. Genome research 14(3): 414-25.MATUKUMALLI, L.; LAWLEY, C.; SCHNABEL, R.; TAYLOR, J.; ALLAN, M.; HEATON, M.; O’CONNELL, J.; MOORE, S.; SMITH, T.; SONSTEGARD, T. y VAN TASSELL, C. 2009. Development and characterization of a high density SNP genotyping assay for cattle. PloS one 4(4): e5350Mac SWEENEY, C. y MACKIE, R. 2012. Micro-organisms and ruminant digestion: state of knowledge, trands and future prospects. Commission On Genetic Resources for Food and Agriculture (Editores), Roma: disponible en: http://www.fao.org/docrep/016/ap099e/ap099e.pdf.METZKER, M. 2010. Sequencing technologies - the next generation. Nature reviews. Genetics 11(1): 31-46.MONIS, P.; GIGLIO, S.; KEEGAN, A. y THOMPSON, R. 2005. Emerging technologies for the detection and genetic characterization of protozoan parasites. Trends Parasitol. 21:340-346.MULLIS, K.; FALOONA, F.; SCHARF, S.; SAIKI, R.; HORN, G. y ERLICH, H. 1986. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. 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