The FISH Assay in Plants

The FISH Assay in Plants

Fluorescence in situ hybridization (FISH) is a valuable cytogenetic technique for the detection and localization of specific DNA or RNA sequences on chromosomes (1). In general, this technique can be used for defining the spatial-temporal expression patterns of target genes. Utilizing fluorescent DNA probes with sequences complementary to targeted chromosomal locations, the gene expression patterns can be easily detected by fluorescent microscope, and further subjected to quantitative analyses (Figure 1). Moreover, by labeling different sequence fragments with distinct fluorophores, the improved FISH assay also allows simultaneous observation of multiple genes (2). Lifeasible provides high quality service for both single- and multi-color FISH assays with improved efficiency and competitive price. Besides, new developed variants of the classical FISH assay have also been designed to largely broaden the application scope of this technique.

FISH.pngFigure 1. A schematic cartoon of the principle of FISH assay

Compared to other in situ hybridization assays, the FISH assay has the following advantages:

  • Enables direct visualization of genetic expression patterns in cells
  • Enables simultaneous examination of multiple genes of interest
  • Quick to perform  
  • High sensitivity and resolution

In the plant system, the FISH assay has been employed in multiple crop species, including wheat, rye, cucumber and melon (3, 4). With solid biochemical and cytological backgrounds, and years of experience in plant cell line/tissue sample generation, scientists and experts here at Lifeasible  provide high quality service that covers every step of the FISH assay in the plant system:

  • Probe designing and construction: aside from commercially available probes, we also provide customized probes that can meet your special needs.
  • Sample preparation: chromosome, cell line and tissue samples
  • Probe/target DNA denaturation and hybridization: our optimized protocol ensures better chromosome spreading, and more efficient hybridization
  • Result acquisition, interpretation and reporting: we and our collaborators have superior microscope system that can obtain high quality images, which allows for  detection of genes even with low copy numbers.

Reference

  1. Langer-Safer PR, Levine M, & Ward DC (1982) Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci U S A 79(14):4381-4385.
  2. Liehr T, Starke H, Weise A, Lehrer H, & Claussen U (2004) Multicolor FISH probe sets and their applications. Histol Histopathol 19(1):229-237.
  3. Fu S, et al. (2015) Oligonucleotide Probes for ND-FISH Analysis to Identify Rye and Wheat Chromosomes. Sci Rep 5:10552.
  4. Shargil D, et al. (2015) Development of a fluorescent in situ hybridization (FISH) technique for visualizing CGMMV in plant tissues. J Virol Methods 223:55-60.
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