Sample 2
Enterococcus faecalis
Protocols: Two cultures of E. faecalis were grown to an OD600 of 0.2 and subsequently split into three to form a total of six cultures (two samples, with three technical replicates each), and grown to an OD600 of 0.5. One set of three cultures were treated with 0.5 ug/ml (0.25 x MIC), and all six cultures were grown for a further 1h. Cells were harvested by centrifugation and stored at -80 degrees C. Cells were harvested by centrifugation in 3:2 glycerol saline at 1Ok x g for 20mins at -10 degrees C. Cells were resuspended in 1:1 glycerol saline, snap frozen and stored at -80 degrees C for RNA extraction the following day. Two 100ml Schott bottles containing 62.5ml of brain heart infusion (BHI) broth were inoculated 1/100 with E. faecalis JH2-2 overnight culture and grown to an OD600 of 0.2 (37 degrees C with gentle agitation). Each culture was split into triplicate and grown to an OD600 of 0.5. One set of triplicates were treated with 0.5ug/ml of teixobactin, while the other set of triplicates were an untreated control. Cells were grown for a further 1 h and harvested as stated above. RNA was extracted by TRIzol-chloroform as per manufacturers instructions. Residual DNA was removed from samples using the Ambion TURBO DNA-free kit. RNA was further purified using the RNeasy Mini Kit (Qiagen) as per the manufacturers instructions. The Illumina TruSeq stranded total RNA library prep kit and ribosomal RNA was removed using Ribo-Zero.
ENA first public
2019-01-31
ENA last update
2018-02-09
External Id
SAMEA104584961
INSDC center alias
University of Otago
INSDC center name
University of Otago
INSDC first public
2019-01-31T17:02:00Z
INSDC last update
2018-02-09T16:21:40Z
INSDC status
public
Submitter Id
E-MTAB-6484:Sample 2
broker name
ArrayExpress
genotype
wild type genotype
growth condition
BHI broth
sample name
E-MTAB-6484:Sample 2
strain
JH2-2