PacBio CCS reads of Emiliania huxleyi virus M1
EhV strains were isolated from water from the induced blooms in mesocosm bags (Schatz et al., 2021) was filtered through a GF/C filter and stored at +4C. This water was used to inoculate E. huxleyi cells and conduct plaque assays for viral isolation. Visible plaques were excised from the agarose at day 4 post-infection and placed in a fresh E. huxleyi CCMP374 culture. Once the culture cleared, the lysate was used for two consecutive plaque assay rounds, confirming the presence of viruses. 100ml cultures of E. huxleyi CCMP374 were grown to exponential phase, then infected with EhVM1. Once the culture cleared, the lysate was filtered (0.45 filter) to eliminate cell debris and the viruses concentrated using a 100kDa Amicon filter. DNA was extracted from the virions by a conventional phenol-chloroform method. Library preparation was according to Pacific Biosciences (PacBio) microbial multiplexing protocol for 1 Sequel small-molecule real-time sequencing (SMRT)-cell (Preparing Multiplexed Microbial Libraries Using SMRTbell Express, Template Prep Kit 2.0, PacBio Biosciences, 2020).