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UDEE 2114 Microbiology Exp1 - Title: Aseptic Technique and...

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Title:Aseptic Technique and Transfer of Microorganism.Objective:To learn the skill of aseptic technique.To isolate the microorganism from a mixed culture to obtain a pure culture.To prevent contamination of microbial cultures and the laboratory or others.Introduction:Aseptic technique is a basic and important skill in the field of microbiology.Microorganism presence in the form of mixed population which aseptic techniqueable to obtain a pure culture by taking care of cleanliness and prevent contaminationof the specific microorganism we are working with. Aseptic technique are series ofprocedures used to reduce the probability of contamination of microbial cultures andprevent contamination of the room and personnel with the microorganism we areworking with.Sterile nutrient-containing-medium which is nutrient agar providing nutrientfor the microorganism to grow. Sterile nutrient agar usually heat to a temperature atwhich all contaminating microorganisms are destroyed. This method is preventing thegrowth of unwanted microorganisms.A pure culture can be obtained through streak plate and pour plate method. Asuccessful growth of pure culture must have sterile agar and apparatuses. Once themicrobial inoculated to the nutrient agar, the colonies should be formed and seenclearly. Colonies actually are formed by a single bacterium which inoculated to theagar and bacterium uses the nutrient to replication and undergoes binary fission. Eachindividual colony showed a single bacterium that has replicated at that location.The sample have transferred to the petri dish which contain nutrient agar willbe growth in a normal temperature (37oC). Most microorganism able to spread andgrow well in this favourable condition.Materials:Nutrient agar, mixed culture, Escherichia coli,Micrococcus luteus,Bacillus subtilis subsp. spizizenii, ethanol at 75% concentration, sterile molten agarApparatus:Inoculating loop, test tube rack, Petri dishes, Bunsen burner, nutrient broth tubes,tissue paper, lighterMethods:Streak Plate1. The table was wiped with 75% concentration of ethanol.2. The Bunsen burner was lighted up.3. The wire loop was flamed under the fire and the top of the petri dish was removedand it was hold by left hand with faced down.4. The sterile loop was dipped into the liquid culture such as (Escherichia coli,Micrococcus luteus) and transferred to the surface of nutrient agar.
5. The culture was transferred to the nutrient agar by moved it gently in a zigzagmanner.6. The loop was removed and flamed in the fire and then allowed it to cool.7. Then, the sterile loop was used to spread the cells enough to get individualcolonies.8. Step 6 and 7 were repeated once again.9. The Petri dish was turned upside down and kept in the favourable environment(37oC) for 1 days.

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Term
Summer
Professor
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Tags
Bacteria, Petri dish, Micrococcus luteus, nutrient agar

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