Sinorhizobium_phage_vB_SmelM_phiM14, T4-like Myovirus infecting Sinorhizobium meliloti 1021
Two separate phiM14 DNA samples, each from a plaque-purified phage sample, were sheared with a Diagenode Bioruptor. Indexed libraries were constructed with an NEBNext Ultra DNA Library Prep kit for Illumina (NEB, Ipswich, MA) in accordance with the manufacturers instructions. For each library, 1 g of DNA was end repaired and ligated to NEBNext adapters. Fragments of an average size 705 bp for sample phiM14.1  were isolated by electrophoresis on Bio-Rad Low Range Ultra Agarose and amplified for 8 to 10 cycles with NEB High-Fidelity 2X master mix and NEBNext multiplex oligonucleotides. Library size distribution was measured on an Agilent Bioanalyzer high-sensitivity chip, and quantity was determined with the KAPA Biosystems Library Quantification kit. Paired-end 300-base sequence reads were generated on an Illumina MiSeq with a 600-cycle MiSeq v3 Reagent kit. Genome assembly from MiSeq reads was performed with Lasergene SeqMan Pro v. 11.2.1.25 (DNAStar, Madison, WI). For both phiM14.1 and phiM14.2 the coverage was approximately 5000x. The initial assemblies predicted for phiM14.1 and phiM14.2 were identical to one another except for two single base mismatches. Both initial assemblies were circularly permuted.