WGS of induced phages from Citromicrobium bathyomarinum strain MCCC1A10434
After fragmented DNA, construct a Library with the NEBNext UltraTM DNA Library Prep Kit for Illumina (NEB, USA) : The End Repair Reaction Buffer (10X) and End Prep Enzyme Mix were added for terminal repair. The reaction procedures were 30min at 20C and 30min at 65C. The NEBNext Adaptor and Blunt/TA Ligase Master Mix were added to the joint and reacted at 20C for 15min. The USER Enzyme was added to remove the base U on the joint and reacted at 37C for 15min. Add 0.35 times the volume of AMPure XP Beads to mix, stand for 5min, place on the magnetic rack for 5min, carefully absorb the supernatant, and then add 0.15 times the volume of AMPure XP Beads for purification. That is, the ligated DNA in the range of 500bp to 700bp is obtained (the inserted fragment is 500bp). Then 6 rounds of PCR amplification were performed. The amplification primers were Universal PCR Primer and Index (X) Primer. PCR reaction conditions were as follows: Pre-denatured 10 Sec at 98C, denatured 10 Sec at 98C, annealed 30 Sec at 65C, extended 30 Sec at 72C; Finally, one-fold AMPure XP Beads were used for purification to obtain sequencable beads