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Page1of8BIOL 211: Introductory MicrobiologyAseptic TechniqueASEPTICTECHNIQUE OFBACTERIALTRANSFERLab 6Studying microorganisms is fundamentally different from the study of larger, macroscopicentities. Because microbes are ubiquitous (present everywhere) and microscopic (invisible to thenaked eye), special procedures must be employed when working withthese tiny creatures.Aseptic techniqueuses the heat from a Bunsenburner to reduce the probability of introducing contaminating culturesduring bacterial transfer from one medium to another. Any time that youtransfer bacterial cultures, you should use aseptic technique. In additionto preventing contamination of the culture being transferred, use ofaseptic technique also ensures that the organism being handled does notcontaminate the handler, others who are present, or the work area.The transfer of bacteria will be conducted using aninoculating loop. This loop must besterilized using the flame of the Bunsen burner before coming into contact with the bacterialsample.BACTERIALCULTURESEach pair of students will share the following cultures at their bench:Escherichia colibrothMicrococcus luteusslantEnterobacter cloacaeagar platebroth culture with mix ofE. coliandM. luteusPROCEDURE(work independently)Work area de-clutter and disinfection1.Remove any items from your bench top that are not necessary for completion of today’sactivities. Personal items can be stored in the cubbies near the sink.2.Use the provided disinfectant and sponge to clean your lab bench.PARTIEach student should obtain:test tube racktwo sterile tryptic soy broth (TSB) tubesone sterile tryptic soy agar (TSA) slantIn the first part of this exercise, you will practice transferring bacteria from one medium toanother using aseptic technique. By the end of Part I, you will complete the following transfers:broth culture (E. coli) to sterile broth tubeagar slant culture (M. luteus) to sterile agar slantagar plate culture (E. cloacae) to sterile broth tubeTransfer of bacteria from broth culture to sterile broth tube1. Label one sterile tryptic soy broth (TSB) tube.Careful: the lid is not water-tight; hold tubeupright.Label should include your name, lab section, lab number, the name of the medium, andaseptic technique:amethod to collect,grow, and preserve asample such that noother microbes areintroduced or lost fromthe original culture
Page2of8BIOL 211: Introductory MicrobiologyAseptic Techniquethe name of bacterium (in this case,E. coli) to be transferred to this tube.2. Hold the handle of the inoculating loop in your dominant handas you would a pencil or chopsticks. Sterilize the inoculatingloop by holding the base of the loop in the flame at a 45°angle.

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