| Journal: | Anais da Academia Brasileira de Ciencias |
| Database: | PERIÓDICA |
| System number: | 000420577 |
| ISSN: | 0001-3765 |
| Authors: | Passamani, Paulo Z1 Carvalho, Carlos R1 Soares, Fernanda A.F1 |
| Institutions: | 1Universidade Federal de Vicosa, Centro de Ciencias Biologicas e da Saude, Vicosa, Minas Gerais. Brasil |
| Year: | 2018 |
| Season: | Mar |
| Volumen: | 90 |
| Number: | 1 |
| Pages: | 41-48 |
| Country: | Brasil |
| Language: | Inglés |
| Document type: | Artículo |
| Approach: | Analítico, descriptivo |
| English 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 |
| Disciplines: | Biología, Química |
| Keyword: | Genética, Bioquímica, Cromosomas, Sondas de ADN, Micromanipulación, Reacción en cadena de la polimerasa (PCR), Hibridación fluorescente in situ (FISH) |
| Keyword: | Genetics, Biochemistry, Chromosomes, DNA probes, Micromanipulation, Polymerase chain reaction (PCR), Fluorescence in situ hybridization (FISH) |
| Full text: | Texto completo (Ver HTML) |
