Cápal Petr, Vrána Jan, Kubaláková Marie, Endo Takashi R, Doležel Jaroslav
Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany, Olomouc, Czech Republic.
Faculty of Agriculture, Department of Plant Life Science, Ryukoku University, 1-5 Yokotani, Seta Oe-cho, Otsu, Shiga, 520-2194, Japan.
Methods Mol Biol. 2016;1469:157-70. doi: 10.1007/978-1-4939-4931-1_12.
Flow cytometry enables chromosomes to be sorted into different groups based on their characteristics, such as relative DNA content and the presence of repetitive DNA sequences. Despite the recent progress in the analysis of plant genome organization and chromosome structure, there is a need for easy methods to assign DNA sequences to individual chromosomes. Here, we describe an easy way to allocate genes or DNA sequences to chromosomes in wheat using flow-sorted chromosomes combined with fluorescence in situ hybridization and PCR analyses.
流式细胞术能够根据染色体的特征,如相对DNA含量和重复DNA序列的存在情况,将染色体分为不同的组。尽管最近在植物基因组组织和染色体结构分析方面取得了进展,但仍需要简便的方法将DNA序列分配到各个染色体上。在此,我们描述了一种利用流式分选的染色体结合荧光原位杂交和PCR分析,将小麦中的基因或DNA序列分配到染色体上的简便方法。