Direct nonradioactive in situ hybridization of somatic cell hybrid DNA to human lymphocyte chromosomes

Kievits, Tim; Devilee, Peter; Wiegant, J.; Wapenaar, Martin C.; Cornelisse, Cees J.; Ommen, G.J.B. van; Pearson, P. L.

doi:10.1002/cyto.990110112

Cited by 52 publications

(19 citation statements)

References 28 publications

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“…1. The first applications of this approach were presented in studies investigating the chromosomal content of interspecies hybrid cells (Boyle et al 1990; Kievits et al 1990). The whole genomic DNA of a hybrid cell line was used as a probe for chromosomal in situ suppression (CISS) hybridization on metaphase spreads from normal cells of a species of interest.…”

Section: Introductionmentioning

confidence: 99%

Detection of amplified DNA sequences by reverse chromosome painting using genomic tumor DNA as probe

et al. 1993

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Abstract.A modification of "reverse chromosome painting" was carried out using genomic DNA from tumor cells as a complex probe for chromosomal in situ suppression hybridization to normal metaphase chromsome spreads. Amplified DNA sequences contained in such probes showed specific signals, revealing the normal chromosome positions from which these sequences were derived, As a model system, genomic DNAs were analyzed from three tumor cell lines with amplification units including the proto

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Section: Introductionmentioning

confidence: 99%

Detection of amplified DNA sequences by reverse chromosome painting using genomic tumor DNA as probe

et al. 1993

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“…Reverse in situ hybridization is an alternative approach to overcome those limitations (9,10). When total hybrid cell DNA is used as a probe against normal human metaphase chromosome spreads, under conditions that suppress signal from ubiquitous repetitive DNA (11)(12)(13)(14)(15), the human DNA present in the hybrid line is specifically delineated.…”

mentioning

confidence: 99%

“…Chromosomal in situ suppression (CISS) hybridization conditions were done as described elsewhere (13 10 ,ug of salmon sperm DNA. Probe denaturation, prehybridizing, and hybridization were done as described (13).…”

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confidence: 99%

Fluorescence in situ hybridization with Alu and L1 polymerase chain reaction probes for rapid characterization of human chromosomes in hybrid cell lines.

Lichter

Ledbetter

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

171

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Human-rodent hybrid cell lines have been analyzed with regard to their human DNA content by using various DNA probe sets, derived from the hybrids, for in situ hybridization to normal human metaphase chromosome spreads. Total genomic hybrid DNA was compared with probe sets of hybrid DNA that were highly enriched in human sequences. The latter probes were obtained by amplification through the polymerase chain reaction (PCR) using oligonucleotide primers directed to human specific subsequences of the interspersed repetitive sequences Alu and Li. Previously unidentified chromosomal material within hybrid lines was characterized with speed and precision. It is demonstrated that the complete human complement of hybrid lines can be rapidly assessed by comparing the data obtained with the Alu-PCR products with the results from the L1-PCR products or from the genomic hybrid DNA. This approach using interspersed repetitive sequence-PCR products is simple and fast and also provides an alternative way of generating complex DNA probe sets for the specific delineation of entire chromosomes or subchromosomal regions by in situ hybridization.Human-rodent somatic cell hybrid lines are very important tools for the analysis of the human genome. In all applications it is crucial to know the complete human chromosomal complement of these lines. The lines are often unstable with regard to chromosomal content and arrangement and, therefore, require periodical re-examinations. Biochemical analysis or Southern blot analysis using large sets of DNA probes is labor-intensive and, in general, an exact determination of the human DNA content is not feasible with these methods. To obtain comprehensive data, cytogenetic analysis is the method of choice. G11 staining of metaphase chromosomes is used to differentiate human from rodent chromosomes on the basis of color (1), but G11 staining is of limited use for the accurate identification of the human chromosomal material.

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“…1986). In situ hybridi zation of biotinylated total hybrid cell DNA to human lymphocyte spreads was performed under repeat suppressing conditions as described by Kievits et al (1990b). Competitive hybridization with biotinylated wholc-cosmid DNA was carried out according to Kievits et al (1990a).…”

Section: Methodsmentioning

confidence: 99%

Isolation and characterization of cell hybrids containing human Xp-chromosome fragments

Wapenaar

Kievits

Khan

et al. 1990

Cytogenet Genome Res

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We have subjected C12D, a Chinese hamster hybrid containing only the human X chromosome, to 6-thioguanine selection. The majority of the derivative clones retained rearranged Xp-fragments, which were characterized by using a combination of enzyme markers, DNA probes, and in situ hybridization. Two of these, TG2 and TG5sc9.1, contained only an Xpter→p21 fragment and should be an ideal resource for directed cloning from this region. A possible mechanism for the specific retention of Xp-fragments is discussed.

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Direct nonradioactive in situ hybridization of somatic cell hybrid DNA to human lymphocyte chromosomes

Cited by 52 publications

References 28 publications

Detection of amplified DNA sequences by reverse chromosome painting using genomic tumor DNA as probe

Detection of amplified DNA sequences by reverse chromosome painting using genomic tumor DNA as probe

Fluorescence in situ hybridization with Alu and L1 polymerase chain reaction probes for rapid characterization of human chromosomes in hybrid cell lines.

Isolation and characterization of cell hybrids containing human Xp-chromosome fragments

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