Protocol for chromosome-specific probe construction using PRINS, micromanipulation and DOP-PCR techniques
An. acad. bras. ciênc
;
90(1): 41-47, Mar. 2018. graf
Artículo
en Inglés
| LILACS
| ID: biblio-886917
ABSTRACT
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.
Texto completo:
Disponible
Índice:
LILACS (Américas)
Asunto principal:
Reacción en Cadena de la Polimerasa
/
Cartilla de ADN
/
Etiquetado in Situ Primed
/
Análisis Citogenético
Tipo de estudio:
Estudios de evaluación
Límite:
Humanos
Idioma:
Inglés
Revista:
An. acad. bras. ciênc
Asunto de la revista:
Ciencia
Año:
2018
Tipo del documento:
Artículo
País de afiliación:
Brasil
Institución/País de afiliación:
Universidade Federal de Viçosa/BR
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