Abstract
BackgroundDevelopment of oligonucleotide probes facilitates chromosome identification via fluorescence in situ hybridization (FISH) in many organisms.ResultsWe report a high throughput and economical method of chromosome identification based on the development of a dye solution containing 2 × saline-sodium citrate (SSC) and oligonucleotide probes. Based on the concentration, staining time, and sequence effects of oligonucleotides, an efficient probe dye of peanut was developed for chromosome identification. To validate the effects of this solution, 200 slides derived from 21 accessions of the cultivated peanut and 30 wild Arachis species were painted to identify Arachis genomes and establish karyotypes. The results showed that one jar of dye could be used to paint 10 chromosome preparations and recycled at least 10 times to efficiently dye more than 100 slides. The A, B, K, F, E, and H genomes showed unique staining karyotype patterns and signal colors.ConclusionsBased on the karyotype patterns of Arachis genomes, we revealed the relationships among the A, B, K, F, E, and H genomes in genus Arachis, and demonstrated the potential for adoption of this oligonucleotide dye solution in practice.
Highlights
Development of oligonucleotide probes facilitates chromosome identification via fluorescence in situ hybridization (FISH) in many organisms
Identification of SSONs for development of the dye solution Four concentrations (2.5 × 10−3 ng/μL; 2.5 × 10−4 ng/ μL; 2.5 × 10−5 ng/μL; and 2.5 × 10−6 ng/μL) of oligonucleotide DP-8 were used to compare signals produced on chromosomes in the dead cell of cultivar Silihong (SLH) with the oligonucleotide dye
The results showed there were no detectable signals at a concentration of 2.5 × 10−6 ng/μL (Fig. 1)
Summary
Development of oligonucleotide probes facilitates chromosome identification via fluorescence in situ hybridization (FISH) in many organisms. Du et al Plant Methods (2019) 15:69 maize [4, 5, 13, 15] This means that many SSONs possibly have the ability to invade double stranded DNA in chromosomes [4]. Du et al [5] found that some SSONs even at very low concentrations, as low as 0.01 ng/μL of (GAA) for example, could still produce clear signals. This is indicative of the efficient length and concentration of some SSONs, which makes it possible to develop a type of SSON dye solution with which chromosomes can be painted
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