Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.12/1442
Título: Heterozygous indels as useful tools in the reconstruction of DNA sequences and in the assessment of ploidy level and genomic constitution of hybrid organisms
Autor: Santos, Carla Sousa
Robalo, Joana Isabel
Collares-Pereira, Maria João
Almada, Vítor Carvalho
Palavras-chave: Double peaks
Interspecific hybrids
Haplotype reconstruction
Beta-actin gene
Cyprinid fish
Data: 2005
Editora: Taylor & Francis
Citação: DNA Sequence, 16 (6), 462-467
Resumo: In this paper we describe a simple approach using double peaks in chromatograms generated as artefacts in the vicinity of heterozygous indels, to identify the specific sequences present in individual strands of a given DNA fragment. This method is useful to assign bases in individuals that are heterozygous at multiple sites. In addition, the relative sizes of the double peaks help to determine the ploidy level and the relative contribution of the parental genomes in hybrids. Our interpretation was confirmed with the analysis of artificial mixtures of DNA of two different species. Results were robust with varying PCR and sequencing conditions. The applicability of this method was demonstrated in hybrids of the Squalius alburnoides complex and in heterozygotes of Chondrostoma oligolepis. Far from being limited to these fish models and the gene where it was tested (betaactin), this sequence reconstruction methodology is expected to have a broader application.
Peer review: yes
URI: http://hdl.handle.net/10400.12/1442
ISSN: 1042-5179
Aparece nas colecções:UIE-E - Artigos em revistas internacionais

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