2005
DOI: 10.1002/0471142905.hg0401s45
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Abstract: Chromosome preparations currently provide the only direct view of the genome as a whole. Although molecular methods allow a more detailed analysis of specific regions of the genome, the study of genetics is not complete without an appreciation of the metaphase cell. The stimulated T cell system described in this unit is the most widely used means of obtaining large numbers of mitotic cells for genetic analyses. Synchronization of the cell cycle in culture is described, combined with direct inhibition of chromo… Show more

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Cited by 20 publications
(8 citation statements)
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References 15 publications
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“…The method used in the present study to prepare chromosome spreads includes incubation with hypotonic [0.075 M] KCl solution. ( 19 ) Because the impairment of condensin subunits leads to weakness against hypotonic shock, the morphological changes observed in the present study, such as ‘blurred’ and ‘amorphous’ chromosome structures, were in agreement with those reported in the condensin subunits‐depleted cells. ( 5,20–22 ) Hudson et al .…”
Section: Discussionsupporting
confidence: 91%
See 1 more Smart Citation
“…The method used in the present study to prepare chromosome spreads includes incubation with hypotonic [0.075 M] KCl solution. ( 19 ) Because the impairment of condensin subunits leads to weakness against hypotonic shock, the morphological changes observed in the present study, such as ‘blurred’ and ‘amorphous’ chromosome structures, were in agreement with those reported in the condensin subunits‐depleted cells. ( 5,20–22 ) Hudson et al .…”
Section: Discussionsupporting
confidence: 91%
“…The cells were incubated with 75 mM KCl in 37°C for 20 min, fixed in 3:1 (v/v) HPLC‐grade absolute methanol/glacial acetic acid, and mixed well gently before spreading onto slides. ( 19 ) Chromosomes staining were carried out in 4% Giemsa solution for 10 min. Cells were observed using an Axioplan‐2 microscope (Carl Zeiss, Oberkochen, Germany), and chromosome lengths (the telomere‐to‐centromere‐to‐telomere length) and widths in cells at prometaphase were measured using IP Laboratory Spectrum v.3.1 (Signal Analytics, Vienna, VA).…”
mentioning
confidence: 99%
“…Cytogenetic examinations were performed on metaphases obtained from peripheral blood lymphocyte cultures using standard procedure [13]. Briefly, the 0.5 mL heparinized peripheral blood samples were cultured at 37°C for 72 h in RPMI-1640 Medium (Sigma-Aldrich, USA), supplemented with fetal bovine serum, (PAN-Biotech, Germany) and Lectins LF-7 (IBSS BIOMED SA, Poland).…”
Section: Methodsmentioning
confidence: 99%
“…Karyotyping was performed as previously described (Bangs and Donlon, 2005), by Karyologic, inc. Briefly, cells were seeded in t-25 tissue culture flasks, and allowed to grow.…”
Section: Methodsmentioning
confidence: 99%