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221 Labplan 2 Isolation Techniques (1) - BIO 221 LABPLAN 2...

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BIO 221 LABPLAN 2 – ISOLATION TECHNIQUESREFERENCESTEXT:Chapter 7.5PPT:Culture MethodsVIDEOS:Aseptic Techniques: Plating Methods– Jove free accessvideoEXERCISE 1:ASEPTIC TRANSFER (WORK INDIVIDUALLY)OBJECTIVEUse aseptic technique to transfer bacteria to solid and broth media.MATERIALSMEDIA:TSB trypticase soy broth (1); TSA trypticase soy agar slant (1)CULTURES:Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus*EQUIPMENT:Inoculating loop, incinerator, labeling tape, test tube rack*Coordinate with students near you so that all four bacteria are representedfor the follow-up lab.PROCEDURE –Each student will set up one of the four bacteria*1.Plug in and turn on the sterilizing incinerator and allow it to heat about 5 minutes.2.Use the same color tape to label a sterile broth and slant with your initials, date,and culture number.Do not write directly on the glass.3.Tighten the cap of the bacterial culture tube and vortex several seconds to mix.4.Insert the wire portion of the loop in the incinerator until it burns bright red.CAUTION: TO AVOID A SERIOUS BURN, NEVER LEAVE THE LOOP UNATTENDED INTHE INCINERATOR.5.Hold the loop 5-10 seconds to allow it to cool, or stand it upright it in the incinerator base or test tuberack (never on the counter – why not?).The loop must be sufficiently cool before touching bacteria toprevent cells from being killed or released into the air as an aerosol.6.Pick up the bacterial culture tube with your free hand.Using the hand that is holding the loop, wrap yourpinky finger around the culture tube cap,turning thetube rather than the cap,to open.The cap shouldremain in your hand with the loop while the tube is open.

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Term
Summer
Professor
NoProfessor
Tags
Bacteria, TSA, tube, inoculating loop, test tube rack, pour plate method

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