Day 1 - Aseptic Transfer of Unknown Bacteria to Prepare Cultures

The beginning of future lab experiments!  In this experiment, we obtained an unknown bacteria from a culture and inoculated it in several other different mediums.  From these cultures, we will isolate our bacteria from the other bacteria that were mixed in the original culture.  Our goal is to identify our isolated bacteria.

Materials:
2 agar slants
1 broth
1 agar plate, with our group number and incubator temperature written on the bottom
Culture of bacteria, kept at 30 degrees Celsius
Bunsen burner
Flint striker
Inoculating loop

Steps:
1.  Light Bunsen burner (Hair and loose clothing should be out of the way)
     a.   Make sure the bottom of the burner is shut tight.
     b.   Turn on the gas
     c.   Carefully loosen the bottom of the burner until gas is heard seeping out
     d.   Ignite the Bunsen burner using a tool given by your professor
     e.   Make the flame low enough that two cones appear
2.  Aseptic transfer of bacteria from sample to agar slant/Slant Inoculation
     a.  Sterilize inoculating loop by holding it in the flame, just above the inner blue cone. (Should turn orange)
     b.  Let the loop cool
     c.  Take cap off of bacterial culture with the same hand holding the loop, flame the opening of the tube, insert sterile loop into culture, withdraw a bead of culture, flame the opening of the tube, replace the cap on the tube, and return the culture to the rack.

Bacteria transfer to broth

     d.  Take cap off of sterile agar slant, flame the opening of the tube, insert inoculating loop into the tube.  Trace the loop on the surface of the agar, squiggling it back and forth like a snake.  Flame the opening of the tube, replace the cap on the tube, and return it to the rack.  Sterilize the inoculating loop. 
     e.  Repeat step "d" for the second agar slant
3.  Aseptic transfer of bacteria from sample to broth/ Broth Inoculation
     a.  Sterilize inoculating loop in the flame
     b.  Let the loop cool
     c.  Take cap off of bacterial culture, flame the opening of the tube, insert sterile loop into culture, withdraw a bead of culture, flame the opening of the tube, replace the cap on the tube, and return the culture to the rack.
     d.  Take cap off of sterile broth, flame the opening of the tube, insert inoculating loop into the tube, swirl the loop around in the broth so the bacteria gets into the culture, flame the opening of the tube, replace the cap on the tube, and return it to the rack.  Sterilize the inoculating loop.
4.  Aseptic transfer of bacteria from sample to petri dish/ Plate Streak
     a.  Sterilize inoculating loop in the flame
     b.  Let the loop cool
     c.  Take cap off of bacterial culture, flame the opening of the tube, insert sterile loop into culture, withdraw a bead of culture, flame the opening of the tube, replace the cap on the tube, and return the culture to the rack.
     d.  Streak one quarter of the petri dish in a back-and-forth motion.  Sterilize the inoculating loop and let cool.  Rotate the petri dish 1/4 of a circle.  Streak the second quarter of the dish.  Sterilize the inoculating loop and let cool.  Rotate the petri dish 1/4 of a circle.  Streak the third quarter of the dish.  Sterilize the inoculating loop and let cool.  Rotate the petri dish 1/4 of a circle.  Streak the fourth quarter of the dish and finish the streak with a little squiggle down the middle of the medium.  Sterilize the inoculating loop.
5.  We placed our cultures in the incubator at 30 degrees Celsius.


Results:  Some good lookin' cultures!

Morphology
Agar plate: circular colonies, cream color, and raised elevation
Agar slant: filiform
Broth: turbid and flocculent