Transformation Lab

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Transformation Lab Student Instructions
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Student Instructions Step 1

• Remove two tubes from the ice bucket.
• Label one B1 and the second "B2."

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Student Instructions Step 2

• Finger flick both tubes to resuspend the cells.
• Open the tube labeled "B1" and use a sterile
  pipette to add one drop of the solution from tube
  "P." Close the tube. Add nothing to tube "B2."

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Student Instructions Step 3

• Place the tubes in ice for 15 minutes.

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Student Instructions Step 4

• Remove the tubes from the ice and immediately
  hold them in a 42 C water bath for 90 seconds.
• Place tubes directly back into the ice bucket for
  1 minute.

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Student Instructions Step 5

• Use a sterile pipette to add 5 drops of sterile
  LB nutrient broth to each of the tubes. Close the
  tubes.

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Student Instructions Step 6

• Allow cell recovery by placing the tubes in a 37
  C water bath for 20-60 minutes.

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Student Instructions Step 7

• Label the underside of the four petri dishes with
  your groups name. On one "Amp" plate, print
  "B1" and on the second "Amp" plate, print "B2."
• Use the same labeling scheme to label the two
  "No Amp" plates.

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Student Instructions Step 8

• Use a fresh sterile pipette to place 3 drops of
  cell suspension from the tube labeled "B1" onto
  the center of a petri dish labeled "Amp/B1" and 3
  drops to the center of a dish labeled "NoAmp/B1."

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Student Instructions Step 9

• Use a second fresh sterile pipette to place 3
  drops of cell suspension from the tube labeled
  "B2" onto the center of a petri dish labeled
  "Amp/B2."

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Student Instructions Step 10

• Use a fresh sterile paper clip to spread the
  liquid evenly across the surface of each plate.
  Do not touch the part of the paper clip that
  comes in contact with the agar.

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Student Instructions Step 11

• You can use one paper clip to spread both plates
  with "B1" solution and one paper clip to spread
  both plates with "B2" solution.

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Student Instructions Step 12

• Incubate the plates (agar down) for 2 hours and
  then invert (agar up) for the next 24-36 hours in
  a 37 C incubator.

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Student Instructions Lab Results
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