Whole genome sequencing of Bacillus cereus bacteriophage phiBc24
Genomic DNA was extracted from the phage particles using a modified phenol-chloroform-isoamyl alcohol method. Briefly, Phage suspension was treated with DNase I and RNase A to remove bacterial DNA contamination. Purified phages were treated with proteinase K, EDTA, and SDS to digest the phage capsid. The DNA phage was purified with an equal volume of phenol/chloroform/isoamyl alcohol (25:24:1) and chloroform/isoamyl alcohol (24:1), then centrifuged. The aqueous layer was collected and an equal volume of isopropanol was added to precipitate the total DNA. The mixture was centrifuged and the DNA pellets were washed twice with ethanol. Lastly, the DNA was dried using Speedvac and resuspended in DNase/Rnase-free deionized water. DNA quality and quantity were determined by NanoDrop 2000 and sequenced by the Illumina HiSeq platform.