Lactobacillus acidophilus for babies,probiotics and human health pdf,is apple cider vinegar like a probiotic - 2016 Feature

In order to better understand what conditions are best suitable for growth of our experimental strain, Bacillus subtilis 168, we tested the growth under various conditions relevant to the work we would perform later. The number and size of colonies which grew on each of the agar plates was recorded as seen below. First we measured the growth curves at various ethanol concentrations to confirm the highest ethanol concentration suitable for cell growth, which was 4%, as displayed below.
Agar plates of varying kanamycin concentration were plated with 100 ?l aliquots of the dilutions, and incubated for ~36 hr at 37° C (Figure 1).


3 ml aliquots of each were put in glass standard bottles, along with varying amounts of kanamycin.
Afterwards, we measured the growth curves at various concentrations of iron, copper, nickel, cobalt, zinc and manganese, which are also displayed below.
This culture was then diluted by a factor of 10:6 and 100ul aliquots were plated onto LB agar plates of varying kanamycin concentrations, and subsequently incubated overnight at 37° C. The number and colour of the colonies were recorded (red indicating presence of pTG262, white being wild type or representing loss of the plasmid before plating).


The Optical density at 600 nm was measured at the start and after 8 hr incubation with shaking at 37° C. It demonstrates both presumed plasmid loss in overnight culture without antibiotic (white colonies) and the decreased growth rate of cells containing pTG262.



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