植物寡核苷酸荧光原位杂交技术方法

Protocals for Oligonucleotide Fluorescence in situ Hybridization in Plants

寡核苷酸荧光原位杂交技术是一种整合了生物信息学分析、DNA高通量合成以及荧光原位杂交实验的新兴技术。该技术适用于任何具有参考基因组的植物物种,并可根据研究需求设计靶向某个染色体区域、整条染色体或一组染色体的寡核苷酸探针,目前已成功应用于数种植物的核型分析、染色体变异、种群演化、同源染色体配对、单倍型分析和异源多倍体识别等研究。该文详述了植物寡核苷酸探针的设计、扩增和标记、染色体制备以及荧光原位杂交的具体操作流程,以期促进寡核苷酸荧光原位杂交技术在细胞遗传学研究中的应用。

Oligonucleotide fluorescence in situ hybridization(Oligo-FISH)is a new technology that integrates bioinfor-matics analysis,high throughput DNA synthesis and fluorescence in situ hybridization experiments.It is applicable to plant species with a reference genome.Oligo probes are able design to a chromosomal region,an entire chromosome,and a set of chromosomes according to researchers’requirements.Oligo-FISH has been successfully applied to studies on karyotyping,chromosome variation,population evolution,homologous chromosome pairing,haplotype analysis,and heteropolyploid identification in several plant species.Here,we introduce the concrete operational processes of oligonu-cleotide-based probe design,amplification and labeling,chromosomes preparation,and fluorescence in situ hybridization in plants,so as to facilitate Oligo-FISH application in the cytogenetics research in the future.