Sexai

Restriction digestions were carried out using the restriction enzymes EcoRI, HincII, NsiI, PstI, SalI, ScaI, SexAI, and SpeI (Thermo Fischer Scientific). These enzymes gave several well separated bands when in silico digested by the NEBcutter (http://nc2.neb.com/NEBcutter2/). The digestions were carried out in a final volume of 10 µL, containing DNA (ca. 300 ng), 0.5 µL of enzyme and 1 µL of Fast digest green buffer (10×, Thermo Fisher Scientific). After 2–16 h incubation at 37 °C the restriction fragments along with undigested DNA and GeneRuler 1 kb DNA Ladder (Thermo Fisher Scientific) were loaded on 1% (w/v) agarose gel including 0.005% (w/v) Midori green. After the electrophoresis the fragments were visualized using UV transillumination and images recorded using the BioRad GelDoc XR+ imaging system.

Restriction digestions were carried out using the restriction enzymes EcoRI, HincII, NsiI, PstI, SalI, ScaI, SexAI, and SpeI (Thermo Fischer Scientific). These enzymes gave several well separated bands when in silico digested by the NEBcutter (http://nc2.neb.com/NEBcutter2/ (accessed on 8 May 2021)). The digestions were carried out in a final volume of 10 µL, containing DNA (ca. 300 ng), 0.5 µL of enzyme, and 1 µL of Fast digest green buffer (10×, Thermo Fisher Scientific). After 2–16 h incubation at 37 °C, the restriction fragments along with undigested DNA and GeneRuler 1 kb plus DNA Ladder (Thermo Fisher Scientific) were separated on 1% (w/v) agarose gel and detected with ethidium bromide staining using UV transillumination. Images were recorded using the BioRad GelDoc XR+ imaging system.