An easy "SteamDrop" method for high quality plant chromosome preparation

The preparation of plant chromosomes for molecular cytogenetic purposes remains a challenge for some species. We studied the dynamics of plant chromosome spreading after dropping cell suspension on slides. We showed that steam (water bath at 55° C) stimulates cytoplasm hydrolysis and rapid chromosome spreading, and that chromosomes stretch during chromosome spreading. Based on these observations, we developed a novel method, named SteamDrop, for preparation of well-spread mitotic and pachytene chromosomes and successfully used it for 28 plant species with large and small chromosomes. We applied cell suspensions in 96% ethanol instead of the commonly used ethanol/acetic acid fixative. Long storage of cell suspensions in ethanol did not impair the quality of chromosome preparations. Mitotic and meiotic chromosomes prepared via SteamDrop were successfully applied for fluorescent in situ hybridization (FISH) with genomic, repetitive and unique DNA probes, as well as for C-banding. The SteamDrop procedure provides a robust and season-independent routine method for high quality chromosome preparations.

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Видео An easy "SteamDrop" method for high quality plant chromosome preparation канала Smelkov Evgeny