Use este identificador para citar ou linkar para este item: https://locus.ufv.br//handle/123456789/16481
Tipo: Artigo
Título: Protocol for chromosome-specific probe construction using PRINS, micromanipulation and DOP-PCR techniques
Autor(es): Passamani, Paulo Z.
Carvalho, Carlos R.
Soares, Fernanda A.F.
Abstract: Chromosome-specific probes have been widely used in molecular cytogenetics, being obtained with different methods. In this study, a reproducible protocol for construction of chromosome-specific probes is proposed which associates in situ amplification (PRINS), micromanipulation and degenerate oligonucleotide-primed PCR (DOP-PCR). Human lymphocyte cultures were used to obtain metaphases from male and female individuals. The chromosomes were amplified via PRINS, and subcentromeric fragments of the X chromosome were microdissected using microneedles coupled to a phase contrast microscope. The fragments were amplified by DOP-PCR and labeled with tetramethyl-rhodamine-5-dUTP. The probes were used in fluorescent in situ hybridization (FISH) procedure to highlight these specific regions in the metaphases. The results show one fluorescent red spot in male and two in female X chromosomes and interphase nuclei.
Palavras-chave: DOP-PCR
FISH
Micromanipulation
PRINS
Editor: Anais da Academia Brasileira de Ciências
Tipo de Acesso: Open Access
URI: http://dx.doi.org/10.1590/0001-3765201720160089
http://www.locus.ufv.br/handle/123456789/16481
Data do documento: 15-Abr-2016
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