Samples were hybridized for 16 hours at 50°C with medium agitation, then sequentially washed in solutions comprising 2× SSC, 0.2% SDS [2×(1 min wash, 1 min soak, 37°C ], 1× SSC [2×(1 min wash, 1 min soak, 37°C ], and 0.5× SSC [2×(1 min wash, 1 min soak, 30°C ], and then dried under N2 (5 min, 30°C).
Strains of S. cerevisiae (LBM-1, CAT-1, and PE-2) and K. marxianus (UFV-3, ATCC 8554, and CCT 4086) were previously grown in 2 % YPD medium under agitation at 180 rpm at 28 °C.
One colony of the bacteria carrying each construct was cultivated at 37°C in 5 mL of Luria Bertani (LB) medium containing ampicillin (100 μg mL-1) (i.e., LB selective medium), under vigorous agitation (200 rpm) until an OD600nm of 0.6 was reached.
The multi-component delivery systems were optimized by evaluating their ability to self-emulsify when introduced to an aqueous medium under gentle agitation.
SEM image of a seeded scaffold incubated in growth medium under constant agitation.
A resuspended P6 pellet derived from 60 ml conditioned medium was incubated for 5 h at 4°C, with 3.5 × 107 magnetic beads carrying an anti-HLA class II monoclonal antibody in 500 μl serum-free culture medium, under mild agitation (Dynabeads-HLA class II, Dynal-Invitrogen).
The first phase was a preselection study employing malt extract medium without agitation.
The tracheae were digested at 4°C overnight in this Digestion Medium without agitation.
Figures 3B and 3C depict the medium volume-agitation rate and initial pH-agitation rate interactions respectively.
Animals were killed by cervical dislocation, and interscapular fat depots were rapidly excised and washed in phosphate-buffered saline, minced and digested for 45 min at 37°C with 2 mg/ml collagenase A (Roche) in the presence of 20 mg/ml bovine serum albumin (Sigma) in Dulbecco's modified Eagle's medium (DMEM) under mild agitation.
