Team:UNC Chapel Hill/30 June 2009



'Frozen Stock' and 'Mini-Prep' in Scott's Lab (6/30 - 7/1)


  • Scott had earlier transformed Ptet mRFP into DH5α-E Chemically competent bacteria. He plated the bacteria and let them culture overnight. Today, Pavel and Peter created culture tubes of this bacteria using the procedure below. They also transformed and plated the LacY gene into the same bacteria.

Procedure to culture cells

  1. Use culture tubes - rounded bottom, 2 level clamp lid. While incubating, remember to close the lid loosely to allow bacteria to get oxygen.
  2. Add 3mL of carb LB to the culture tube.
  3. Take a clean pipette tip greater than the 3mL level in the tube, touching only the very top of it.
  4. Carefully scoop up a single colony from the agar dish with the pipette tip and drop it into the culture tube. (Once again, make sure the part that you touched does not go into the LB. Use a fairly large pipette tip.)
  5. Label the side of the tube and put into the shaking incubator at 37 degrees C overnight.

Altered Procedure from June 4th

  1. Used LB (Luria Broth: 10g Trypton, 5g yeast extract, 10g NaCl, 1L water) instead of SOC as the growth medium.
  2. Only used 3 Microcentrifuge Tubes with CC cells.