RAD-seq of Scutellastra granularis pool
We used the ezRAD library preparation and sequencing approach (Toonen et al. 2013), which uses high-frequency cutter isochimozer enzymes MboI and Sau3AI (which cut the same site) to perform a DNA double digest. Digestion and library preparation protocols were the same of those described in Toonen et al. (2013). Briefly, the digested DNA is inserted into an Illumina TruSeq library preparation kit, where the digesting libraries are end-repaired and TruSeq adapters are ligated to the genomic fragments. Libraries are then size-selected on a 2100 Agilent Bioanalzer, quantified using qPCR (following Kapabiosystems Illumina quantification kit protocol), and paired end sequencing using 600 cycles on the Illumina Mi-Seq platform. Ê