Protocol for chromosome-specific probe construction using PRINS, micromanipulation and DOP-PCR techniques

dc.creatorPassamani, Paulo Z.
dc.creatorCarvalho, Carlos R.
dc.creatorSoares, Fernanda A.F.
dc.date2018-01-18T13:37:45Z
dc.date2018-01-18T13:37:45Z
dc.date2016-04-15
dc.date.accessioned2023-09-27T20:56:05Z
dc.date.available2023-09-27T20:56:05Z
dc.identifier1678-2690
dc.identifierhttp://dx.doi.org/10.1590/0001-3765201720160089
dc.identifierhttp://www.locus.ufv.br/handle/123456789/16481
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8951177
dc.descriptionChromosome-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.
dc.formatpdf
dc.formatapplication/pdf
dc.languageeng
dc.publisherAnais da Academia Brasileira de Ciências
dc.relationDec. 2017
dc.rightsOpen Access
dc.subjectDOP-PCR
dc.subjectFISH
dc.subjectMicromanipulation
dc.subjectPRINS
dc.titleProtocol for chromosome-specific probe construction using PRINS, micromanipulation and DOP-PCR techniques
dc.typeArtigo


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