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Titolo:
RAPID DETECTION OF CHROMOSOMES BY IN-SITU HYBRIDIZATION OF LABELED OLIGONUCLEOTIDES AND COMPARISON WITH PRINS
Autore:
COULLIN P; VALENT A; BARBOUNAKI I; CANDELIER JJ; PELLESTOR F; BERNHEIM A;
Indirizzi:
INSERM,U178,16 AVE PAUL VAILLANT COUTURIER F-94807 VILLEJUIF FRANCE INST GUSTAVE ROUSSY,CNRS UPR 9008 F-94805 VILLEJUIF FRANCE
Titolo Testata:
Comptes rendus de l'Academie des sciences. Serie 3, Sciences de la vie
fascicolo: 10, volume: 319, anno: 1996,
pagine: 901 - 906
SICI:
0764-4469(1996)319:10<901:RDOCBI>2.0.ZU;2-I
Fonte:
ISI
Lingua:
FRE
Soggetto:
INSITU HYBRIDIZATION;
Keywords:
MOLECULAR CYTOGENETICS; LABELED OLIGONUCLEOTIDES; IN SITU HYBRIDIZATION; PRINS; CHROMOSOME 1;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
7
Recensione:
Indirizzi per estratti:
Citazione:
P. Coullin et al., "RAPID DETECTION OF CHROMOSOMES BY IN-SITU HYBRIDIZATION OF LABELED OLIGONUCLEOTIDES AND COMPARISON WITH PRINS", Comptes rendus de l'Academie des sciences. Serie 3, Sciences de la vie, 319(10), 1996, pp. 901-906

Abstract

We propose a simple, fast and inexpensive method of identification ofhuman centromeres on metaphasic chromosomes and interphasic nuclei. This is based on in situ hybridization of labelled oligonucleotides. The efficiency of the methodology was demonstrated on cytogenetic preparations from human heteroploid and human x hamster hybrid cell lines and also on frozen tissue sections using an oligonucleotide specific forthe alpha-satellite DNA of chromosome 1. Three versions of this oligonucleotide respectively labelled with 1, 4 and 10 fluorescein molecules were synthesized. The signal intensity provided by the oligonucleotide coupled with 4 fluoresceins allowed unambiguously the detection of the chromosome and the establishment of its ploidy using a classical cytogenetic microscope without the need for an amplification procedure. The use of different fluorochromes and possibly combination with an unlabelled elongation in 3' of the oligonucleotides which stabilize itshybridization, lead to a simple multicolour method. Preliminary quantification of the signals obtained by in situ hybridization of labelledoligonucleotides and comparison with those obtained by primed in situlabelling (PRINS) using the same nucleotides as primers, suggest thatthe elongation generated by PRINS may be very short compared with a PCR in solution. This limited efficiency of the in situ elongation may reflect the present difficulties of PRINS and DISC PCR (direct in situsingle copy polymerase chain reaction) with primers specific for non-repetitive sequencies.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 26/09/20 alle ore 07:00:10