WGS of Pluralibacter gergoviae UPMC1178
The expressed breast milk was collected from 30 healthy woman who were breastfeeding children from birth to two years old. The samples were received as freshly expressed, chilled or iced. All samples collected were immediately stored at -20 C until further analyses. Isolate was obtained by plating 100 L of the milk sample on non-selective MRS agar plates after thawing at 4 C overnight. The plates were incubated aerobically at 37 C for 48 to 72 h. An MRS medium was used to support growth of nutritionally fastidious microbes. Based on colony morphology, colonies were randomly picked from culture plates and purified twice in the same conditions for 18 to 24 h. The purified strains were stored at -80C in growth media containing 20% (v/v) glycerol. It was then subculture into LB broth. The isolate was cultured into a total of 8 plates. All the plates were incubated for 16 hours. Next, one single colony was transferred to 5ml LB broth and was incubated for overnight, shaked for 180rpm. DNA extraction was performed using DNA extraction Kit (Qiagen, USA). The extracted DNA was sent to Codon Genomics Sdn. Bhd for genome sequencing. The sequencing was conducted by using HiSeq 4000 (Illumina, US).
Pluralibacter gergoviae UPMC1178