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A combined microsatellite multiplexing and boiling DNA extraction method for high-throughput parentage analyses in the Pacific oyster (Crassostrea gigas) ArchiMer
Taris, Nicolas; Baron, S; Sharbel, Tim; Sauvage, Christopher; Boudry, Pierre.
The analysis of parentage using microsatellite markers is of increasing importance, notably in aquaculture genetic research where communal rearing of mixed families can be used to reduce unwanted environmental variance. We present here an optimization of parental genotype assessment for larvae or adults of the Pacific oyster, Crassostrea gigas, using a multiplex system of three microsatellite loci. In conjunction with a simple DNA extraction protocol, this method enables high throughput analyses of parentage in C. gigas. Using this method, we successfully determined the parentage of 93% (1224 out of 1318) of the progeny in a factorial cross between 3 females and 10 males. The inability to genotype the remaining 7% was due to DNA degradation of larvae...
Tipo: Text Palavras-chave: Oysters; Aquaculture; Parental assignment; High throughput; Larval DNA extraction; Multiplex.
Ano: 2005 URL: http://archimer.ifremer.fr/doc/2005/publication-1814.pdf
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A multiplex PCR assay able to simultaneously detect Torque teno virus isolates from phylogenetic groups 1 to 5 BJMBR
Devalle,S.; Niel,C..
Torque teno virus (TTV) is a circular, single-stranded DNA virus that chronically infects healthy individuals of all ages worldwide. TTV has an extreme genetic heterogeneity which is reflected in its current classification into five main phylogenetic groups (1-5). Using specific PCR assays, it has been shown that many individuals are co-infected with TTV isolates belonging to different phylogenetic groups. Here, a multiplex PCR assay was developed, using five recombinant plasmids. Each plasmid carried an insert of different size issued from a TTV isolate belonging to a different group. The assay was able to simultaneously amplify DNAs of TTV isolates belonging to all five phylogenetic groups. Multiplex PCR was then tested satisfactorily on DNAs extracted...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Brazil; Co-infections; Multiplex; PCR; Torque teno virus.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2005000600006
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Duplex nested-PCR for detection of small ruminant lentiviruses BJM
Marinho,Rebeca C.; Martins,Gabrielle R.; Souza,Kelma C.; Sousa,Ana Lídia M.; Silva,Sabrina Tainah C.; Nobre,Juliana A.; Teixeira,Maria F.S..
Abstract Small ruminant lentiviruses (SRLV) have high genetic variability which results in different viral strains around the world. This create a challenge to design sensible primers for molecular diagnosis in different regions. This work proposes a protocol of duplex nested-PCR for the precise diagnosis of SRLV. The technique was designed and tested with the control strains CAEV Co and MVV 1514. Then, field strains were submitted to the same protocol of duplex nested-PCR. Blood samples of sheep and goats were tested with AGID and nested PCR with specific primers for pol, gag and LTR. The AGID results showed low detection capacity of positive animals, while the nested PCR demonstrated a greater capacity of virus detection. Results demonstrated that...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Standardization; Multiplex; Viruses; CAEV; MVV.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000500083
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