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ABSTRACT
This project examines a detailed process taken in the isolation of plasmids from the rhizobium of Groundnut plant. Plasmid are usually circular molecules of DNA, They can only multiply inside a host cell. Most plasmids inhabit bacteria. Plasmids are also found in higher organisms such as yeast and fungi. The Groundnut (Arachis hypogea) also known as peanuts is a legume crop grown mainly for its edible seeds was procured from the NEW BENIN market in Benin city and planted in a cross experiment with beans in a number of 36 rubber containers containing loam soil. The plant was monitored and taken care of for a time period of five weeks as they grew to a length of at least 6 inches. They were then uprooted and the rhizobium attached to the root nodules of the groundnut host plant was harvested, washed and infused into different Eppendorf tubes for transportation to the lab. Upon arrival in a sterile lab with the rhizobium, they were taken out of the Eppendorf tubes, then washed with hypo solution then placed in beakers to be crushed with a sterilized pipette handle to expose the plasmids of the rhizobium. The crushed rhizobium was then kept in a broth medium in conical flasks for storage until the next day to grow the bacteria, a YEMA (Yeast Extract Mannitol Agar) medium was prepared and poured in multiple amounts of petri-dishes and kept for a day to solidify. A toothpick was used to streak the rhizobium in the broth solution across the YEMA medium in a lab sterilized by ethanol flame and covered to facilitate the growth of the rhizobium on the YEMA medium in the petri dish. After the growth of the rhizobium is noticeable to the naked eye, a bit of cut off and was made to undergo gram staining technique which allowed us to know it was a gram-negative-bacteria. The rhizobium bacteria already prepared in the petri-dish is then made to undergo plasmid DNA extraction by following various instructions required in alkaline lysis of plasmid DNA.
