Question

Pseudomonas putia is thought to degrade hydrocarbons due to the fact that it harbours a TOL plasmid. To verify this assertion, a student was tasked to isolate and grow this soil-dwelling bacteria and to also isolate the plasmid from the microbe. The TOL plasmid is also intended to be used as a cloning vector.
Demonstrate how you could isolate and culture Pseudomonas putida from soil sample contaminated with hydrocarbons.

Answer 1

We can do it by 2 grams of homogenized hydrocaebon contaminated soil sample is added to 50ml of enrichment medium containing salts and peptone water, yeast extract,some hydrocarbons like hexadecane.now cultures are grown in 250ml Erlenmeyer flask and incubated with shaking at 150 rpm for 7 days at 37 degree Celsius and at 30 degree Celsius for purpose of isolating pseudomomas pudita.then after some time the culture is placed in solid medium salt media with hexadecane for culture and growth is seen for the bacteria colonies gets formed after some days of pseudomomas pudita which can be taken out of culture by inoculation loop and you have the bacteria separated.