Genetic Engineering: Bacterial Transformation Lab

In this lab we are see in if any genetic transformation is taking place when we place bacteria into different situations. We left them to sit overnight to get a result in the morning. My group found this lab to be very interesting. Down below are the instructions, data collections and a few pictures of the after results.

INSTRUCTIONS:

1. Label one closed micro test tube +pGLO and another -pGLO. Label both tubes with your group's name. Place them in the foam tube rack.
2. Open the tubed and using a sterile transfer pipet, transfer 250 ul of transformation solution (CaC12).
3. Place the tubes on ice.
4. Use  sterile loop to pick up a single colony of bacteria from your starter plate. Pick up the +pGLO tube and immerse the loop into the transformation solution at the bottom of the tube. Spin the loop between your index finger and thumb until the entire colony is dispersed in the transformation solution (with no floating chunks). Place the tube back in the tube rack in the ice. Using a new sterile loop, repeat for the -pGLO tube.
5. Examine the pGLO plasmid DNA solution with the UV lamp. Note your observations. Immerse a new sterile loop into the plasmid DNA stock tube. Withdraw a lapful. There should be a film of plasmid solution across the ring. This is similar to seeing a soapy film across a ring for blowing soap bubbles. Mix the lapful into the cell suspension of the =pGLO tube. Close the tube and return it to the rack on ice. Also close the -pGLO tube. Do not add plasmid DNA to the -pGLO tube.
6. Incubate the tubes on ice for 10 minutes. Make sure to push the tubes all the way down in the rack so the bottom of the tubes stick out and make contact with the ice.
7. While the tubes are sitting on ice, label your 4 agar plates on the bottom (not the lid) as follows: LB/amp plate: =pGLO; LB/amp/ara plate: +pGLO; LB/amp plate: -pGLO; Label the LB plate: -pGLO
8. Heat shock. Using the foam rack as a holder, transfer both the (+) pGLO and (-)pGLO tubes into the water bath, set at 42C, for exactly 50 seconds. When the 50 seconds are done, place both tubes back on ice.
9. Remove the rack containing the tubes from the ice and place on the bench top. Open a tube and using a new sterile pipet, add 250 ul of LB nutrient broth to the tube recluse it. Repeat with a new sterile pipet for the other tube. Incubate the tubes for 10 minutes at room temperature.
10. Tap the closed tubes with your finger to mix. Using a new sterile pipet 100 ul of the transformation and control suspensions onto the appropriate plates.
11. Use a new sterile loop for each plate. Spread the suspensions evenly around the surface of the afar by quickly skating the flat surface of a new sterile loop back and forth across the plate surface.

DATA COLLECTION:

1. Carefully draw and observe what you see on each of the four plate.    (Pictures are listed below)
2. How much bacterial growth do you see on each plate, relatively speaking?  There is a lot of growth. It looks like there are thousands of tiny spots.
3. What color are the bacteria?  The bacteria is a tannish, whitish-yellow color.
4. How many bacterial colonies are on each plate (count the spots you see).  To many to count.