Strategy: WGS
Source: GENOMIC
Selection: RANDOM
Layout: PAIRED, Orientation: , Nominal length: 150, Nominal Std Dev: 30
Construction protocol: A large culture (400 ml) in late log phase growth was harvested by centrifugation, washed in 0.9 g/l NaCl solution, and suspended for cell wall digestion by EDTA and lysozyme, followed first by proteinase K digestion for 60 min at 65 *C, then RNAse digestion (at 37 deg C overnight) to eliminate RNA. The suspension was subjected to phenol extraction to remove proteins and repeated phenol-chloroform extractions before ethanol precipitation. The nucleic acid fraction was centrifuged, and the pellet dissolved in TE buffer, pH 8. The protocol and conditions followed those of Sambrook J., and D.W. Russell. 2001. Molecular Cloning: a laboratory manual. 3rd ed..