AI Article Synopsis
- A new fluorescence hybridization (FISH) protocol was created for nematodes that uses electroporation to introduce nucleic acid probes directly into the organism.
- This streamlined method reduces chemical wash steps, leading to lower specimen loss and faster protocol times, specifically optimized for juveniles of soybean cyst nematodes.
- The protocol allows for the simultaneous detection of two different targets using long wavelength fluorophores, which enhances hybridization sensitivity and may be adapted for other nematode species and life stages.
Article Abstract
A fluorescence hybridization (FISH) protocol was developed for nematodes in which nucleic acid probes are introduced within the organism electroporation. This modification of existing FISH protocols removes numerous chemical wash steps, and thus, reduces protocol time and specimen loss while improving hybridization sensitivity. The presented work is optimized for juveniles of soybean cyst nematode (SCN; ) and has been used to identify both host and associated-microbial (viral) targets. Moreover, through the use of two different long wavelength fluorophores, two probes can be colocalized within one individual. This protocol may be adapted to identify targets-of-interest within other life stages and nematode species. This protocol improves: •Hands-on protocol time (by approximately 1.5 h).•Specimen loss (fewer aspiration steps).•Hybridization (allows colocalization with two nucleic acid probes and increases sensitivity).
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Source |
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6881680 | PMC |
| http://dx.doi.org/10.1016/j.mex.2019.11.009 | DOI Listing |
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