Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720-3220, USA.
Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA.
G3 (Bethesda). 2022 Jul 6;12(7). doi: 10.1093/g3journal/jkac121.
Repetitive DNA sequences are useful targets for chromosomal fluorescence in situ hybridization. We analyzed recent genome assemblies of Caenorhabditis elegans and Pristionchus pacificus to identify tandem repeats with a unique genomic localization. Based on these findings, we designed and validated sets of oligonucleotide probes for each species targeting at least 1 locus per chromosome. These probes yielded reliable fluorescent signals in different tissues and can easily be combined with the immunolocalization of cellular proteins. Synthesis and labeling of these probes are highly cost-effective and require no hands-on labor. The methods presented here can be easily applied in other model and nonmodel organisms with a sequenced genome.
重复 DNA 序列是染色体荧光原位杂交的有用目标。我们分析了秀丽隐杆线虫和太平洋真涡虫的最新基因组组装，以鉴定具有独特基因组定位的串联重复序列。基于这些发现，我们为每个物种设计并验证了至少针对每个染色体 1 个基因座的寡核苷酸探针集。这些探针在不同组织中产生可靠的荧光信号，并且可以很容易地与细胞蛋白的免疫定位相结合。这些探针的合成和标记具有很高的成本效益，不需要人工操作。本文介绍的方法可以很容易地应用于其他具有测序基因组的模式生物和非模式生物。
