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After shake-off or mild trypsinization, mitotic cells were lysed in 75 mM KCl. Chromosomes were spread on slides in the Cytospin (2,000 rpm for 10 min).
Polytene chromosomes were spread on adhesion microscopic slides (Menzel SuperFrost® Plus) by iterative dropping from 20 to 30 cm height.
The chromosomes were spread according to the method described by Shoguchi et al. and stained with Giemsa.
Chromosomes were spread on dry, acetone-cleaned slides by applying drops of cell suspension from a height of 15 cm to 20 cm.
For some experiments polytene chromosomes were spread onto microscopic slides: whole cells were gently separated ~48 hours post conjugation using a Heraeus Megafuge at 1000 rpm, 4°C, 7 min. Cells were resuspended in methanol:acetic acid (3:1) and incubated overnight.
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Since chromosomes are spread out in these structures, the spindle pool of dynein was more separated from the chromosomal pool, allowing visualization of dynein cupping the bivalent ends.
It is also interesting that the sequences from the different Pan troglodytes chromosomes are spread between the different blocks of similarity, suggesting they may have resulted from the duplication of a block of sequences from one chromosome onto another, rather than the tandem duplication of a sequence once it has been transferred to that chromosome.
In general, Y-chromosomes might be spread via a process of demic diffusion during the early agricultural expansion period [65], [66].
A similar survey in zebrafish found that, although there is a cluster of Class III genes on chromosome 19, Class III loci are spread across all chromosomes [ 8].
For example, on chromosome VIII, 18 markers are spread on the top 71 cM while 167 markers are spread on the next 29 cM.
Although chromosomes 2 and 6 contain four and three of the 12 heat shock genes analyzed, respectively, the remaining four are spread among chromosomes 3, 4 and 7 (Fig. 1).
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