1. Aseptic Technique: Media and Equipment

2. Growth Medium
A growth medium or culture medium is a liquid or gel designed to _______ ____ _______ of microorganisms or cells
The most common growth media for microorganisms are ________ _______ and _______ plates

3. Defined Media
An important distinction between growth media types is that of ________ versus __________ media
A defined medium will have _________ quantities of all ingredients

4. Undefined Media
An undefined medium has some complex ingredients that consist of a mixture of many chemical species in _________ _____________
Undefined media are sometimes chosen based on _______ and sometimes by ____________ - some microorganisms have never been cultured on defined media.

5. Undefined Media We will use this in the lab Nutrient Agar contains:
________ _______ ___ ________
________ ________
________

6. Forms of Culture
The most common growth media for microorganisms are ________ _______(liquid in a test tube)
Liquid media are often mixed with ______ and poured into ______ _______ to solidify
These agar plates provide a _______ medium on which microbes may be cultured. They remain solid, as very few bacteria are able to decompose agar.

9. Slants There are several reasons a slant is best for culture storage:
1) ________ ______ – slant increases it
2) ______ – test tubes smaller to store
3) ________ – won’t dry out
4) _______ – once the microorganism is transferred, you can store it for up to 6 months in a refrigerator

11. Aseptic Technique
Aseptic technique refers to a procedure that is performed under ________ ____________
This includes ________ and _________ techniques (such as with microbiological cultures)
It includes techniques like
_________ ______________
(Bunsen burner)

12. Aseptic Technique
Sterile surfaces must be protected from ________ ___ ____ ____ or on non-sterile surfaces
In sterile technique, only ________ surfaces touch other ________ surfaces and ____ exposure is kept to a minimum

13. Aseptic Technique
It is important in microbiology to work with ________ __________
This is difficult because the world around us is covered with ________ (even on dust particles in the air)
In order to protect broth, plates, slants and pure cultures from MO’s around us, we practice _________ __________

14. Aseptic Technique
In this class you will need to practice sterile technique when we _________ a pure culture into ______ __________ (tube of sterile broth or an agar plate)

15. Aseptic Technique “A” = Negative prefix “Septic” = Infection
All techniques and procedures which __________ _______________

16. Safe Patterns Safe patterns include: Standard _______ _________
______________ Techniques
______ and ________ of Equipment
_________ Hands

17. Safe Patterns
________ all materials before beginning

18. Safe Patterns
Spray the lab top down with a _________ _________ or a ______ _______ __________ and allow this to stand for a _______. You may then wipe down the bench with the paper towel.

19. Safe Patterns
Wash hands _______ and _______ lab.

20. Safe Patterns
You should have only the _________ ______ ________ and the written _____ __________ on your bench top or desk.

21. Safe Patterns
Keep petri dishes and test tubes ___________ as much as possible.
If top must be removed completely do ______ _____ _____on the lab top. This lowers the probability of contamination and prevents “false positive” results.

22. Safe Patterns
Hold bottles and tubes ____ ___ _______ to minimize the amount of airborne microbes that can fall into them (blue circle). Remove the caps as shown above and ___ _____set the caps down. Keep the mouth of the cap facing _________ (red circle).

23. Safe Patterns
When using metal ________ ________, HEAT the _______ piece of metal of the inoculation instrument in the flame: it should be RED HOT. Be sure to ________ your inoculation instrument ________ picking the inoculum (broth or agar).

24. Safe Patterns
To __________ a Petri plate: Lift one edge of the Petri plate cover to gain access to the culture medium. Keep the cover over the plate bottom to prevent ______ and __________ from falling onto the agar.

25. Safe Patterns Report _________ to me immediately
Cover the spill with ______ ________ and squirt _________ onto the towels. Wait _____ _________ then clean up the spill.

26. Safe Patterns
________ all test tubes and petri plates with your name (initials), date, and name of organism ________ you add any solutions, bacteria, etc. 

27. Safe Patterns
Do _______ dump ANY microbial suspension down the ______ or in the _______ ______. I will collect them for proper disposal.

28. Safe Patterns Place _____ ________in racks when working at your table:
never lay the tubes down—____ _____
Keep test tube ______ and petri dish _______ on media to reduce _______________ (matters not whether it is sterile media or already cultured).

29. Safe Patterns All agar plates are incubated _______ ______
to reduce bacterial
_______________ and to
reduce the possibility of water
_______________ that may be on the lid
dropping onto the agar, causing fluid to
run across the agar medium.  

30. Media
__________ after preparation, in storage, or working containers. Handle _____________.
If no goofs – Sterile ___________

31. E. Autoclave _____________ ________
Water boils at 100ºC, but if pressure is __________ the boiling point _______
At 15 PSI the Boiling Point = 121ºC
At 20 PSI the Boiling Point = 126ºC
Minimum = ____ ____ PSI
A pressure cooker is the same as an ____________

32. Boiling
_________ kills _______ MO, fungi, and viruses but a few are ____________.
Example: Inf. Hepatitis, Endospores
But very good nonprofessional use

33. Technique depends on article:
Equipment
Technique depends on article:
Does it _____ or _____?
Do you want it back?

34. Equipment Autoclave If it _____ into machine and ____ ____
___ 121ºC (15 PSI) minimum

35. Equipment
Boiling
________ in a lab

36. Equipment Oven _____ ______ used for metal and glass
____ _________ minimum

37. Equipment
___________ – Burning
________ Plus
Loops, test tubes

38. Equipment _____________ Equipment Usually _______ and ______ materials
Gas: _______ _______
____________