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Lab 1: Aseptic Technique and Isolation of Bacteria

Lab 1: Aseptic Technique and Isolation of Bacteria. Terms used in Aseptic Technique. Aseptic Technique : procedure to prevent contamination of medium or bench surface. Pure culture : contains only 1 bacterium Mixed culture : 2 or more species living together

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Lab 1: Aseptic Technique and Isolation of Bacteria

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  1. Lab 1: Aseptic Technique and Isolation of Bacteria

  2. Terms used in Aseptic Technique • Aseptic Technique: procedure to prevent contamination of medium or bench surface. • Pure culture: contains only 1 bacterium • Mixed culture: 2 or more species living together • Inoculation: act of placing bacteria (and other microorganisms) onto culture medium. • Contaminant: unwanted microbes present in culture medium or lab bench surface. • Sterile Media: media prepared and then sterilized prior to use. • Always inspect media to ensure no visible contaminants are present prior to use. Media may be sterilized by autoclaving or filtration.

  3. Important Terms for Aseptic Procedure Inoculating Loop (a)/Needle (b): metal wire used to transfer organisms Incinerator: heat source that is used to any unwanted microorganisms on the inoculating loop/needle.

  4. Media Types and Uses • Broth: a liquid medium. Advantage: tube is easy to store and transport. Disadvantage: can not see colony morphology. • Slant: tube of solid medium at an angle. Advantage: tube is easy to store and transport, can see colony morphology. Disadvantage: small surface area. • Agar deep: tube of solid or semi-solid medium. Good for organisms that prefer reduced O2 and to evaluate motility. Broth Slant Agar deep

  5. Media Types and Uses • Petri plate: solid medium on flat surface. This is the best method to observe colony morphology and to work with individual colonies. Best for diagnostic methods. Petri Plate

  6. Removing inoculum from broth

  7. Removing inoculum from a solid medium

  8. Inoculating Bacteria on Agar Slant Do NOTgouge the agar with the inoculating loop, instead gently graze the surface.

  9. Inoculating Bacteria in Deep Agar Stab the needle containing bacteria directly into the deep agar.

  10. Microorganisms to be used this semester Many of the microorganisms we will use this semester will be Biosafety Level 1 (not shown to cause disease in humans) but several will be Biosafety Level 2 (can cause disease in humans). Because of this potential risk we ask that you treat ALL bacterial cultures as if they cause infection!

  11. Aseptic Technique Lab: Part 1 The detailed procedure can be found on pages 7-8

  12. Inoculating Bacteria on Agar Plate:The Streak Plate Technique

  13. Inoculating Bacteria on Agar Plate:The Streak Plate Technique

  14. Inoculating Bacteria on Agar Plate:The Streak Plate Technique

  15. Improper Streak Plating Technique

  16. Streak Plate Technique Lab: Part 1

  17. Assignments for this week Lab reports – Aseptic Technique & Isolation Streak Plate • Answer all questions

  18. Patterns of Growth in Broth

  19. Patterns of Growth on Agar Slants

  20. Patterns of Growth in Deep Agar

  21. Patterns of Growth on Agar Plate

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