RNAseq of Macrostomum lignano DV1_10 subline: adult F1 offspring from pool C, replicate 1
We established two new sublines, DV1_8 and DV1_10, with the karyotypes 2n=8 and 2n=10, respectively, differing with the copy number of the largest chromosome MLI1. Each subline was initiated from two karyotyped individuals of the inbred line DV1 and cultivated for six months (~8 generations) under standard laboratory conditions. We generated five pools of worms from each subline, DV1_8A-E and DV1_10A-E; each pool was initiated from 20 juvenile worms. Their offspring F1 was regularly collected and cultivated separately until their maturation. F1 generation of each pool was characterized by karyotyping of 40 specimens. Total RNA extracted from pooled worms (N = 60 for each sample) using PureZOL (Bio-Rad, ) according to the manufacturers instructions. The samples were treated with DNase I (New England Biolabs, USA) to eliminate DNA contamination. The total RNA was purified on Agencourt RNAClean XP beads (Beckman Coulter, Germany). Directional RNA-seq libraries were prepared in accordance with New England Biolab protocols. Briefly, polyA-tailed mRNA was purified from 300 ng of total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, USA, E7490S). Then, directional cDNA libraries were generated by means of the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs, USA) following the manufacturers recommendations, and index codes were added to attribute sequences to each sample. The libraries were sequenced on an Illumina Illumina NovaSeq 6000 platform.