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Registro Completo |
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Biblioteca(s): |
Embrapa Mandioca e Fruticultura. |
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Data corrente: |
15/04/2004 |
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Data da última atualização: |
15/04/2004 |
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Autoria: |
MBA, R.; TOHME, J.; ROCA, V.; FREGENE, M. |
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Título: |
Lowering the cost of biotechnologies for national cassava programs: microsatellite markers to facilitate use of the cassava molecular genetic map and provide new genetic information. |
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Ano de publicação: |
1998 |
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Fonte/Imprenta: |
Revista Brasileira de Mandioca, Cruz das Almas, v. 17, p. 25, nov., 1998. Suplemento. |
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Idioma: |
Inglês |
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Conteúdo: |
Abstract: The applicabillity of the majority of molecular markers, currently available on the molecular genetic map of cassava, in marker-assisted studies of complex traits and marker-assisted improvement of cassava has been questioned by collaborators in the NARs considering the low technology status of most cassava breeding program. The speed and low cost of PCR markers. with high levels of allelic diversity. easy to assay. and codominant make them the markers of choice for transfer of gene tags for important cassava traits to collaborators in the National Programs. PCR technology requires a relatively small capital out-lay for its integration into day-to-day plant breeding operations and it eliminates labor costs involved in the tedious Southern analysis of RFLP markers. More microsatellite markers can be assayed for a large number of plants, in a given time, for a given budgest, compared to RFLP markers. A DNA library, enriched for microsatellite markers. was developed by the affinity capture method and cloned into pUC 18 plasmid. The library was transformed into DH 10B competent cells. A total of 18.000 colonies from the enriched library was screened for microsatellite markers by hybridization with microsatellite oligonucleotides and by anchored primer PCR. A total of 120 positive clones were identified and sequenced on an AB1377 automated sequencer. Primers. 20-mers long. were designed from regions flanking microsatellite markers and were used in PCR amplification of the parental genotype of the mapping population. Polymorphic microsatellite markers were scored in the progeny and mapped onto the existing map of cassava using the linkage analysis software MAPMAKER 3.0 Results reveal a rather low efficiency of enrichment using the affinitv capture method, though different di and tri-nucleotide microsatellite markers could be obtained. Approximately 200 mapped microsatellite markers is the targest of this ongoing project. MenosAbstract: The applicabillity of the majority of molecular markers, currently available on the molecular genetic map of cassava, in marker-assisted studies of complex traits and marker-assisted improvement of cassava has been questioned by collaborators in the NARs considering the low technology status of most cassava breeding program. The speed and low cost of PCR markers. with high levels of allelic diversity. easy to assay. and codominant make them the markers of choice for transfer of gene tags for important cassava traits to collaborators in the National Programs. PCR technology requires a relatively small capital out-lay for its integration into day-to-day plant breeding operations and it eliminates labor costs involved in the tedious Southern analysis of RFLP markers. More microsatellite markers can be assayed for a large number of plants, in a given time, for a given budgest, compared to RFLP markers. A DNA library, enriched for microsatellite markers. was developed by the affinity capture method and cloned into pUC 18 plasmid. The library was transformed into DH 10B competent cells. A total of 18.000 colonies from the enriched library was screened for microsatellite markers by hybridization with microsatellite oligonucleotides and by anchored primer PCR. A total of 120 positive clones were identified and sequenced on an AB1377 automated sequencer. Primers. 20-mers long. were designed from regions flanking microsatellite markers and were used in PCR amplification of ... Mostrar Tudo |
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Categoria do assunto: |
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Marc: |
LEADER 02523naa a2200157 a 4500
001 1651862
005 2004-04-15
008 1998 bl uuuu u00u1 u #d
100 1 $aMBA, R.
245 $aLowering the cost of biotechnologies for national cassava programs$bmicrosatellite markers to facilitate use of the cassava molecular genetic map and provide new genetic information.
260 $c1998
520 $aAbstract: The applicabillity of the majority of molecular markers, currently available on the molecular genetic map of cassava, in marker-assisted studies of complex traits and marker-assisted improvement of cassava has been questioned by collaborators in the NARs considering the low technology status of most cassava breeding program. The speed and low cost of PCR markers. with high levels of allelic diversity. easy to assay. and codominant make them the markers of choice for transfer of gene tags for important cassava traits to collaborators in the National Programs. PCR technology requires a relatively small capital out-lay for its integration into day-to-day plant breeding operations and it eliminates labor costs involved in the tedious Southern analysis of RFLP markers. More microsatellite markers can be assayed for a large number of plants, in a given time, for a given budgest, compared to RFLP markers. A DNA library, enriched for microsatellite markers. was developed by the affinity capture method and cloned into pUC 18 plasmid. The library was transformed into DH 10B competent cells. A total of 18.000 colonies from the enriched library was screened for microsatellite markers by hybridization with microsatellite oligonucleotides and by anchored primer PCR. A total of 120 positive clones were identified and sequenced on an AB1377 automated sequencer. Primers. 20-mers long. were designed from regions flanking microsatellite markers and were used in PCR amplification of the parental genotype of the mapping population. Polymorphic microsatellite markers were scored in the progeny and mapped onto the existing map of cassava using the linkage analysis software MAPMAKER 3.0 Results reveal a rather low efficiency of enrichment using the affinitv capture method, though different di and tri-nucleotide microsatellite markers could be obtained. Approximately 200 mapped microsatellite markers is the targest of this ongoing project.
700 1 $aTOHME, J.
700 1 $aROCA, V.
700 1 $aFREGENE, M.
773 $tRevista Brasileira de Mandioca, Cruz das Almas$gv. 17, p. 25, nov., 1998. Suplemento.
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Registro original: |
Embrapa Mandioca e Fruticultura (CNPMF) |
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