Melinaea marsaeus DNAseq PacBio sequences
DNA extraction extracted following this Mugford and colleagues protocol (Mugford et al. 2020) with the following modifications: frozen tissue (approx. 100 mg) was ground in a 2 ml Eppendorf containing a 2 mm steel ball bearing in a Retsch mill (Qiagen), and extracted in 500 l CTAB buffer (2% w/v CTAB, 1.4 M NaCl, 20 mM EDTA, 100 mM Tris-HCl pH 8.0) with RNAse A (20g/ml), incubated at room temperature for 1 hour with regular mixing. 500 l of chloroform was adde d, mixed by inversion for 10 minutes, and centrifuged at 5000 g for 10 minutes. The upper phase was transferred to a new tube, an equal volume of cold isopropanol was added, and incubated at -20 degrees C for one hour. The sample was centrifuged at 5000g for 10 minutes, and the supernatant removed. The pellet was washed in 75% ethanol air-dried, and resuspended in 80 l of TE (10 mM Tris-HCl pH 7.5, 1 mM EDTA). Library construction including adaptor ligation and size selection were performed according to the manufacturers instructions. The PacBio library was sequenced on 10 SMRTcells of the PacBio RSII with C4-P6 chemistry.