Consulta: creatorFacets:"David, E"
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Mark-recapture cloning: a straightforward and cost-effective cloning method for population genetics of single-copy nuclear DNA sequences in diploids
Provedor de dados: 5
Autores: Bierne, Nicolas; Tanguy, A; Faure, Michel; Faure, B; David, E; Boutet, I; Boon, E; Quere, N; Plouviez, S; Kemppainen, P; Jollivet, D; Moraga, D; Boudry, Pierre; David, P.
We describe a simple protocol to reduce the number of cloning reactions of nuclear DNA sequences in population genetic studies of diploid organisms. Cloning is a necessary step to obtain correct haplotypes in such organisms, and, while traditional methods are efficient at cloning together many genes of a single individual, population geneticists rather need to clone the same locus in many individuals. Our method consists of marking individual sequences during the polymerase chain reaction (PCR) using 5'-tailed primers with small polynucleotide tags. PCR products are mixed together before the cloning reaction and clones are sequenced with universal plasmid primers. The individual from which a sequence comes from is identified by the tag sequences upstream...
Tipo: Text
Palavras-chave: Sequence; Population genetics; High throughput allele recognition; DNA polymorphism.
Ano: 2007
URL: http://archimer.ifremer.fr/doc/2007/publication-4222.pdf