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- Title
Oligonucleotide Fluorescence In Situ Hybridization: An Efficient Chromosome Painting Method in Plants.
- Authors
Harun, Arrashid; Liu, Hui; Song, Shipeng; Asghar, Sumeera; Wen, Xiaopeng; Fang, Zhongming; Chen, Chunli
- Abstract
Fluorescence in situ hybridization (FISH) is an indispensable technique for studying chromosomes in plants. However, traditional FISH methods, such as BAC, rDNA, tandem repeats, and distributed repetitive sequence probe-based FISH, have certain limitations, including difficulties in probe synthesis, low sensitivity, cross-hybridization, and limited resolution. In contrast, oligo-based FISH represents a more efficient method for chromosomal studies in plants. Oligo probes are computationally designed and synthesized for any plant species with a sequenced genome and are suitable for single and repetitive DNA sequences, entire chromosomes, or chromosomal segments. Furthermore, oligo probes used in the FISH experiment provide high specificity, resolution, and multiplexing. Moreover, oligo probes made from one species are applicable for studying other genetically and taxonomically related species whose genome has not been sequenced yet, facilitating molecular cytogenetic studies of non-model plants. However, there are some limitations of oligo probes that should be considered, such as requiring prior knowledge of the probe design process and FISH signal issues with shorter probes of background noises during oligo-FISH experiments. This review comprehensively discusses de novo oligo probe synthesis with more focus on single-copy DNA sequences, preparation, improvement, and factors that affect oligo-FISH efficiency. Furthermore, this review highlights recent applications of oligo-FISH in a wide range of plant chromosomal studies.
- Subjects
PAINTING techniques; PLANT hybridization; CHROMOSOMES; FLUORESCENCE in situ hybridization; TANDEM repeats; PLANT chromosomes; FLUORESCENCE
- Publication
Plants (2223-7747), 2023, Vol 12, Issue 15, p2816
- ISSN
2223-7747
- Publication type
Article
- DOI
10.3390/plants12152816