BI 352L: medical micro protocols & terms

Heating time of an inoculating loop/needle

10 seconds
Mouth of glassware – 3s
Glass slides (for heat fixation) – 1

Heating time of mouth of glassware

3 seconds

Heating of glass slides

10 seconds

What is heating glass slides used for?

heat fixation

What kinds of liquid medias will we be using?

tryptic soy broth (for growing liquid cultures)

durham tube (looking at fermentation of carbs, captures gas, upside down)

What kind of agar will we be using?

petri plates with agar on the bottom, agar side up for storage, single colonies can form if not

Where does an orange colony on a stored plate go?

orange colony goes in biohazard bin, bacteria growing on plates stored

What kinds of solid media will we be using?

slant, solid agar to not move, to grow bacteria on slant for stocks of cultures

simon citrate, type of slant,

agar deeps, agar is upright ont slanted, agar on bottom for motility of bacteria

What is aseptic technique’s purpose?

Preventing contamination of cultures

How is sterile handling used in the lab?

• wipe hands & work with 3% hydrogen peroxide/70% ethanol

• wear gloves

• don’t pour media/reagent directly from bottles/flasks

• use sterile glass/disposable plastic pipettes

• never uncover sterile flas/bottle/petri dish until use and cover quickly

• put caps with the opening down

• use only sterile glassware & equipment

• don’t talk, sing, whistle in sterile procedures

• do experiments quickly

T or F: we demonstrated incorrect aseptic technique to demonstrate importance of aseptic technique

True

T or F: it is unnecessary to flame the mouth of the stock culture when inoculation a new broth culture

false, should always be flamed

T or F: you should heat an inoculating loop in the incinerator broth before & after use.

True, to ensure it is sterile

T or F: you should flame the mouth of bottles of test tubes for 3s.

True, mouths are flamed for 3s

T or F: when inoculating agar in a petri dish, before you begin you should remove the lid of the plate and set it on the bench.

False, allows air to contaminate.