Chromosome Classification and Purification Using Flow Cytometry and Sorting

Gray, J. W.; Langlois, Richard G.

doi:10.1146/annurev.bb.15.060186.001211

Cited by 63 publications

(40 citation statements)

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“…The success of carrying out FISH on sorted chromosome would provide a unique starting material that enables physical mapping genome. Flow-sorted chromosomes have been effectively exploited in human chromosomes (Gray and Langlois 1986). The sorted chromosomes of a single type from plants have also been used to construct chromosome-specific library and physically map DNA sequences at the chromosomal level (Arumuganathan et al 1994;Macas et al 1996;Li et al 2004).…”

Section: Discussionmentioning

confidence: 99%

“…Flow-sorted chromosomes have been effectively exploited for human chromosomes (Gray and Langlois 1986). Flow-sorting of chromosomes could be done from various plant species including Haplopappus gracilis (De Laat and Schel 1986), Petunia hybrida (Conia et al 1987), Lycopersicon pennellii (Arumuganathan et al 1994), Vicia faba (Macas et al 1996), Hordeum vulgare (Lysak et al 1999;Lee et al 2000), Melandrium album (Kejnovsky et al 2001), Triticum aestivum (Gill et al 1999;Kubalakova et al 2002), Pisum sativum (Neumann et al 2002), Secale cereale (Kubalakova et al 2003) and Zea may (Li et al 2004).…”

Section: Introductionmentioning

confidence: 99%

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Flow-Sorted chromosomes: a fine material for plant gene physical mapping

Kevin³

et al. 2006

Caryologia

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-Fluorescence in situ hybridization (FISH) on flow-sorted plant chromosomes is described. The protocol involves flow cytometric sorting of metaphase chromosomes, then fixing them with 4% paraformaldehyde solution, and re-sorting these chromosomes directly onto a spot on poly-lysine coated slides after stained. FISH mapping experiments were performed on the sorted chromosomes from oat and barley using 45S and 5S ribosomal DNAs as probes. The chromosome morphology was very well preserved and there were nearly no chromosomes lost after procedures of hybridization and fluorescent staining. Sorted chromosomes are advantageous over metaphase chromosomes as targets for FISH mapping studies because a large number of target chromosomes with better chromosome morphology on a small area on the slide are easy to gain by flow-sorting, background is very clear and hybridization sensitivity is enhanced. The successful and reliable location of rDNA genes in these two plant genomes, combining with the data published before in maize proves that flow-sorted chromosomes are fine materials for gene physical mapping in plants.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Flow-Sorted chromosomes: a fine material for plant gene physical mapping

Kevin³

et al. 2006

Caryologia

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“…Each distribution was derived from a specific gated region of the hamster flow karyotype which contained selected, well-isolated human chromosome peaks. The human chromsome locations in these distributions were determined in other studies (5). Table 3 compares (11).…”

Section: Human/hamster Hybrid Studiesmentioning

confidence: 99%

Centromeric index versus DNA content flow karyotypes of human chromosomes measured by means of slit‐scan flow cytometry

Lucas

Gray

1987

Cytometry

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We have applied slit-scan flow cytometry (SSFCM) to classify human chromosomes according to their centromeric index (CI) and relative DNA content. The resulting bivariate-CI vs. DNA content-distributions shows 14 peaks for normal human chromosomes. Distinct peaks are produced by chromosomes 1, 2, 3, 4+5, 6+7+X, 8, 13+14+15, 16, 17+18, 19+20, and 21+22+Y. In addition, chromosomes 9 through 12 are resolved into three peaks. The identity of the chromosomes comprising each peak was determined by comparing CT vs. DNA content distributions measured for normal human chromosomes by means of SSFCM with CI and DNA content values measured for human chromosomes with image analysis. The accuracy of CI measurement by SSFCM was verified by measuring CIS for human chromosomes isolated from humadrodent hybrid cell lines containing only a few known human chromosomes. These studies showed CIS measured for human chromosomes 1-19 and 21 to be in close agreement with the CIS calculated by means of image analysis. We further confirmed the chromosome assignments for each peak by showing that the relative volumes of the peaks in the CI vs. DNA content distributions for chromosomes from normal cells are similar to the relative frequencies of chromosomes expected for these peaks based on the peak assignments.

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“…While considerable progress has been made concerning the automated evaluation of dicentric chromosomes from homogeneously stained specimen (Gray and Langlois, 1986;Lorch and Stephan, 1986), the automated detection of other structural aberrations, such as translocations, is still in the beginning (Piper and Lundsteen, 1987;Willborn et al, 1987).…”

Section: Introductionmentioning

confidence: 99%