Bacterialglycerolstocks To 2mls of mid-log culture or 1ml of freshly saturated culture add 1 ml(or an equal volume) of glycerol solution, mix gently, then freeze rapidly in liquid nitrogen or on dry ice. Store at -20°C and - 70°C. Glycerol solution: (Per 100 ml) 65% glycerol (v...
For several experiments, inoculation was performed by diluting bacterial glycerol stocks stored at −80°C in a 1:100 ratio into fresh brain heart infusion (BHI) medium (Oxoid, Basingstoke, UK). Neutrophil isolation. Neutrophils were freshly isolated from four healthy donors (two females age ...
Glycerol stocks were stored at −80 °C. To activate cultures, bacteria were passaged at least three times. Bacteria were cultured on their respective agar plate types, temperatures, and oxygen conditions in a confluent cell lawn (Supplemental Table S2). Anaerobic conditions were achieved by ...
S. typhimuriumSL7207 strains expressing scrambled-shRNA and cMyc-shRNA were revived from glycerol stocks 12 h before scheduled co-culture. A scraping of a glycerol stock was placed in 5 ml of LB broth with kanamycin (50 μg/ml) and grown in a shaking incubator at 37 °C for 8 h. 3...
Finally, the Elution buffer (25mM Tris-HCl pH = 8.0, 150mM NaCl, 10% Glycerol, 0.1% NP40, 500mM Imidazole) was used to eluted the recombinant protein. Amicon Ultra centrifugal filters (10kD) were used to concentrate the proteins and exchange buffer (25 mM HEPES pH 7.5, 250 mM ...
(ii) transformants were recovered in SOC medium for 6 h at 37 °C and patched onto LB/agar plates supplemented with 50 μg/ml Amp, 25 μg/ml Kan, 25 μg/ml Spc, and 15 μg/ml Chl; (iii) plates were incubated at 37 °C for 48 h; (iv) glycerol stocks ...
solanacearum strains for tomato, pepper and eggplant were cultured from glycerol stocks from −80 °C lab glycerol stocks and used in this experiment. Three treatments were set up for each crop: WRS (water with pathogen), PRS (10 mM prebiotics with pathogen), and NPRS (10 mM non-...
other nasty critter. If the plates are old, either transform a new set of cells or chip of a bit of frozen glycerol stock from the top of the tube with a pipet tip (do NOT let the frozen cell thaw) and spread on an LB Agar plate with antibiotic. Culture overnight in the 37 ...
When cooled, inoculate the Hungate tubes with 1 mL of the glycerol stocks Place the tubes in a static 37° C. incubator for about 16 hours. The following day, take 1 mL of this subculture and inoculate 10 mL of YCFA (pre-warmed flushed Hungate tubes again, all in duplicate) Place the...
When cooled, inoculate the Hungate tubes with 1 mL of the glycerol stocks Place the tubes in a static 37° C. incubator for about 16 hours. The following day, take 1 mL of this subculture and inoculate 10 mL of YCFA (pre-warmed flushed Hungate tubes again, all in duplicate) Place the...