Answer:
Explanation:
The objective here is to prepare 1 liter of LB + Kan (100 μg/ml final concentration) + Amp (50 μg /ml final concentration) liquid media.
Given that :
the Stock concentration of Amp: 100 mg/ml (since 1 mg/ml = 1000 μg/ml)
it is required that we convert stock concentration in μg/ml since 100 mg/ml = 100000 μg/ml
However, using formula C₁V₁=C₂V₂ (Ampicilin),
where:
C₁ = 100000 μg/ml,
V₁=?,
C₂= 50 μg/ml,
V₂=1000 ml
100000 μg/ml × V₁ = 50 μg/ml × 1000 ml
V₁ = 50 μg/ml × 1000 ml/100000 μg/ml
V₁ = 0.5 ml
Given that:
the Stock concentration of Kan: 25 mg/ml (since 1 mg/ml = 1000 μg/ml)
it is required that we convert stock concentration in μg/ml , 25 mg/ml = 25000 μg/ml
Now by using formula C₁V₁=C₂V₂ (Kanamycin),
C₁ = 25000 μg/ml,
V₁=?,
C₂= 100 μg/ml,
V₂=1000 ml
25000 μg/ml × V₁ = 100 μg/ml × 1000 ml
V₁ = 100 μg/ml × 1000 ml/25000 μg/ml
V₁ = 4 ml
Thus; in 1 lite of Lb+ Kan+Amp preparation;
0.5 ml of Amp & 4 ml of kanamycin is used for their stock preparation.
Finally;
Sterilization step should be carried out in flask (Clean dry glass wares) for media in an autoclave, the container size should be twice the volume of media which is prepared.
