Preliminary work demonstrated that extraction of entire cultures (media and cells together) resulted in the same level of extraction efficiency as cultures that were separated by centrifugation in glass tubes into cell pellet and media followed by separate extraction of the separated cell pellet and media.
To estimate the use and the yield of the extraction of blood cultures in an Intensive Care Unit (ICU) and to assess the variables associated with the yield.
For extraction of metabolites cultures were harvested by filtration, washed with ddH2O, and quenched in liquid nitrogen.
A HS extraction of the culture broth of A. camphorata followed by incubation on a carboxen/polydimethylsiloxane (CAR/PDMS) fibre during 31.8 min at 54.6 °C gave the most effective and accurate extraction of the volatile compounds.
A close approximation would involve direct quenching and extraction of the culture broth (i.e. cells + supernatants, therefore removing the problematic washing step) and a differential/subtractive method for estimation of intracellular metabolite concentrations [31].
Total blue pigments levels were determined by extraction of whole culture samples with 1 M KOH (final concentration) and spectrophotometric measurement at 640 nm [ 27].
For DNA extraction, two loopfuls of cultures grown on PDA agar (Oxoid Ltd., Basingstoke, Hampshire, England) for 2 to 5 days at 25°C were suspended in 500 μL of lysing buffer (50 mM Tris, 250 mM NaCl, 50 mM EDTA, 0.3% sodium dodecyl sulfate (SDS) (pH 8.0)) plus the equivalent of a 200 μL volume of 425 to 600 um diameter glass beads (Sigma).
DNA extraction of the pure cultures, 16S rRNA gene amplification, and sequencing were performed according to [ 18].
The first evidence is that the extraction of biosurfactant from culture medium is not feasible both technically and economically; therefore, the raw biosurfactant in culture broth could be used directly for the injection process (Rabiei et al. 2013; Sarafzadeh et al. 2013; Souayeh et al. 2014).
NH and HK carried out the RNA extraction of the primary culture, cell line (MCF-7) and biopsy specimens.
