ddRAD of Porcellio spinicornis: adult pereion and leg tissues
Library preparation followed the ddRADseq approach of Peterson et al. (2012) and was performed by the UAlberta Molecular Biology Service Unit. Preparation followed MacDonald (2020) with the following modifications: (1) the restriction enzymes were EcoR1 and Pst1; (2) all sixteen i5 adaptors were used, but subpools were made with 10 microlitres of eight sample ligations instead of 5 microlitres of 16 samples (step 3); (3) purification of the final pool was done with a 1:1 ratio of Seramag Select beads following the manufacturer's protocol; 4) size range of the final pool was 400-2000 (average 1000 bp). Sequencing was performed using a NextSeq 500 (Illumina) and a 75-cycle high output kit. Demultiplexing was performed in the process_radtags program in Stacks v2.64 (Rochette et al. 2019). Reads were demultiplexed and filtered for quality with Stacks v 2.64 (Rochette et al. 2019) with further filtering in fastP v 0.23.1 (Chen et al. 2018) to remove enzyme cut sites and Illumina adaptor sequences. Final cleaned sequences are 63 bp long.
SUB15660909
090AAA