Genomic DNA extraction
Genomic DNA was extracted from samples containing a pool of fish eggs
and larvae fixed in ethanol. To ensure the complete evaporation of the
alcohol, initially the excess was removed through pipetting, and then
the microtubes were kept open for three hours at 55°C. We then added
600µl of TNES buffer to each sample and ground the bulk with a plastic
pestle until only minuscule tissue fragments were left. Next, 20µl of
proteinase K (20mg/ml) was added to each microtube. The samples were
kept at 55°C until complete tissue digestion. Finally, the genomic DNA
was extracted using a low-cost saline protocol adapted from (Aljanabi
and Martinez, 1997).
The samples were quantified using a Qubit 4 Fluorometer (Thermo Fisher)
with a 1x dsDNA HS Assay Kit (Thermo Fisher) to verify the success of
the DNA extraction. The samples were then diluted to 100ng/µl.