There is a significant difference in viscosity (p < 0.05) between NE formulations prepared with different poloxamer types and the same concentrations (weight by weight percent "w/w %") of oil, surfactant mixture and co-surfactant.
The second random PCR (50 μl) contained 5 μl of the first PCR products as template and the same concentrations of the other constituents, and the reaction was run for 30 cycles under the same conditions.
The second random PCR mixture (50 μl volume) contained 1 μl of the first PCR product as template and the same concentrations of constituents used in the original reaction.
After the adsorption period, the inoculum was removed, the cells washed once with PBS and the same concentrations of each extract and fractions diluted in culture media without SFB were added to complete 24 h at 37 °C in a 5%% CO2 atmosphere.
Mixture B contained 0.5 mM ATP, 0.5 mM GTP, 4 µM [H]fMet-tRNAfMet, 4 µM IF2 and the same concentrations of PEP and Mg OAc 2 in polymix buffer as in mixture A. Mixture A was pre-incubated for at least 10 min at 37°C to ensure ribosome dissociation into 30S and 50S subunits.
Mixture B contained 2 µM 50S ribosomal subunits, 0.5 mM ATP, 0.5 mM GTP and the same concentrations of PEP and Mg OAc 2 in polymix buffer as in mixture A. The formation of 70S initiation complexes was then initiated by mixing mixtures A and B in a stopped-flow instrument as described above.
