HMW gDNA for building reference genome for Rhizoglyphus robini
eggs were smashed with a sterile pellet pestle on ice in 500ul homogenization buffer (HB); the sample was fixed with 500ul cold ethanol 60 minutes on ice, after that time the sample was centrifuged at 1500 g for 5 minutes at 4C, and the supernatant was discarded. Resuspended in HB pellet was cast in four agarose plugs as described in the original protocol. Agarose plugs were incubated with Proteinase K and Lysis buffer solution for 2h with intermittent mixing. After that time digestion solution was replaced with a freshly made one and incubated overnight with intermittent mixing. According to the original protocol, after RNase A digestion and plug washing, DNA was recovered by incubation of the plugs in TE buffer, followed by plug melting and addition of agarase. Recovered DNA was dialyzedand homogenized on a membrane for 45 minutes at room temperature, and transferred to a clean tube with a wide bore tip.