ArticleBiology open2026
A protocol for sequential isolation, purification, and collection of Luticola nematophagus eggs to obtain high-quality genomic DNA.
Article in Biology open, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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11 authors.
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Abstract
Conventional strategies for obtaining planarian tissue for genomic DNA extraction rely on diet-based rearing or amputation-induced regeneration. These approaches are inapplicable to soil-dwelling flatworms that predominantly reproduce by egg-laying and lack regenerative capacity, such as the recently discovered nematode predator Luticola nematophagus. Here, we developed a protocol for the sequential isolation, purification, and collection of L. nematophagus eggs, comprising three steps: (1) physical sieving through sintered meshes of graded pore sizes under recirculating water flow to exclude nontarget eggs; (2) chemical digestion with 8% (w/w, weight/weight) urea or 5% (w/w) hydroxylamine hydrochloride under continuous aeration for 18 h to eliminate contaminating microinvertebrates; and (3) manual sorting under a stereomicroscope. Genomic DNA extracted from purified eggs exhibited A260/A280 ratios of 1.93-2.00 and A260/A230 ratios exceeding 2.0, meeting the quality thresholds for whole-genome sequencing. Successful PCR amplification of 18S and 28S ribosomal DNA (rDNA) fragments confirmed the suitability of the DNA for downstream molecular analyses. Compared with untreated controls, urea treatment (8% urea and manual sorting) reduced the final hatching rate by 5.2% and delayed the half-hatching time by 5.1 days, whereas hydroxylamine hydrochloride treatment (5% hydroxylamine hydrochloride and manual sorting) reduced the final hatching rate by 11.6% and delayed the half-hatching time by 7.9 days. However, the majority of the eggs remained viable. The quality of the extracted DNA was further confirmed by successful whole-genome sequencing, which yielded a chromosome-level genome assembly, indicating its suitability for the most demanding downstream applications.
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