 Nutrients - C, H, O, N, S. P, K, Mg, Fe, Ca, Mn, and traces of Zn, Co, Cu, and Mo. These elements are found in the form of water, inorganic ions, small molecules, and macromolecules which serve either a structural or functional role in the cells.  Energy source- All living organisms require a source of energy. Organisms that use radiant energy (light) are called phototrophs. Organisms that use (oxidize) an organic form of carbon are called heterotrophs or (chemo)heterotrophs. Organisms that oxidize inorganic compounds are called lithotrophs.  Growth Factors- amino acids, nucleic acids and vitamins  Optimum temperature- 37 degree celsius or 98.6 degree fahrenheit

 Strict/Obligate Aerobes- require oxygen  Facultative anaerobe- can survive with or without oxygen  Obligate/Strict Anaerobe- die in the presence of oxygen  Microaerophile- require lower levels of oxygen to survive.

 Introduction of bacteria into various forms of culture media in order to keep them alive and to study their growth. Contamination should be avoided at all cost during that process.

 Broth- Liquid media without agar. Most bacterial samples can be easily introduced into and grown in this type of medium, even those with widely different aero tolerance. Inoculating loop used.  Slant – Solid media in test tube kept in a slanted position. Inoculating loop used.  Deep – Semi- Solid media( % agar) in test- tube kept in upright position. Needle is used.  Plate – Solid media grown in plates to provide a large surface area

Composition:  0.5% PeptonePeptone  0.3% beef extract/yeast extractbeef extractyeast extract  1.5% agaragar  0.5% NaClNaCl  distilled water distilled water  pH adjusted to neutral (6.8) at 25 °C.

Two types of seaweed, Gelidium, and Gracilaria, are harvested to produce agar. Agar actually consists of two substances, called agarose, and agaropectin. The microbes feed off the added nutrients, but cannot digest the agar, so it remains intact, allowing colonies of organisms to be easily observed and studied. Agar is a gel at room temperature, remaining firm at temperature as high as 65°C. Agar melts at approximately 100°C, a different temperature from that at which it solidifies, °C.

 Solid- % of agar is 1-3%(Generally 1.5%)  Semi-Solid- % of agar is %

 Used to separate/isolate different organisms from one another by diluting the original concentration of the sample in which they were contained.  Isolating a pure culture is important to identify an unknown microorganism

 Pour Plate  Streak Plate  Spread Plate

 the purpose of streaking for isolation is to produce isolated colonies of an organism on an agar plate.isolated colonies  This is useful when you need to separate organisms in a mixed culture or when you need to study the colony morphology of an organism.

 Flame your loop after streaking each quadrant.  Cool the loop before touching agar  Do not blow in order to cool the loop  Keep the plate covered partially while streaking  Do the streaking near the flame

 YOUR NAME/Lab partner’s name  LAB SECTION  NAME OF SPECIMEN  DATE Use only water proof ink pens. NO PENCILS. Masking tape is used to label the test tubes.

 Clean the working area with disinfectant  Wash your hands  Wear your lab coat. Write your name on the lab coat.  Take 1 inoculating needle,1 loop,1striker and 1 wax pencil from the drawers.  Please tie your hair to protect it from fire/ bacteria

 Keep the nutrient agar plates (INVERT) and the tubes in incubator  Turn off the gas nozzle.  Wash your hands and disinfect the working area.  Keep your lab coats back in drawer.  Collect your pre-labs, review and return it to me.