Faculty of Biology, Institute of Botany, Dresden, Germany.
Methods Mol Biol. 2023;2672:151-162. doi: 10.1007/978-1-0716-3226-0_8.
The quality of chromosome preparation influences all downstream analyses and is therefore crucial. Hence, numerous protocols exist to produce microscopic slides with mitotic chromosomes. Nevertheless, due to the high content of fibers in and around a plant cell, preparation of plant chromosomes is still far from trivial and needs to be fine-tuned for each species and tissue type. Here, we outline the "dropping method," a straightforward and efficient protocol to prepare multiple slides with uniform quality from a single chromosome preparation. In this method, nuclei are extracted and cleaned to produce a nuclei suspension. In a drop-by-drop manner, this suspension is then applied from a certain height onto the slides, causing the nuclei to rupture and the chromosomes to spread. Due to the physical forces that accompany the dropping and spreading process, this method is best suited for species with small- to medium-sized chromosomes.
染色体制备的质量会影响所有下游分析，因此至关重要。因此，存在许多方案可以产生具有有丝分裂染色体的显微镜载玻片。然而，由于植物细胞内外纤维含量高，植物染色体的制备仍然远非易事，需要针对每种物种和组织类型进行微调。在这里，我们概述了“滴注法”，这是一种从单个染色体制备中制备具有均匀质量的多个载玻片的简单而有效的方案。在该方法中，提取和清洁核以产生核悬浮液。以逐滴的方式，将该悬浮液从一定高度滴到载玻片上，使核破裂并使染色体展开。由于滴注和展开过程中伴随的物理力，该方法最适合中小染色体的物种。
