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Protocol for chromosome-specific probe construction using PRINS, micromanipulation and DOP-PCR techniques Anais da ABC (AABC)
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...
Tipo: Info:eu-repo/semantics/article Palavras-chave: DOP-PCR; FISH; Micromanipulation; PRINS.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652018000100041
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