User:Hana Benzeer/Notebook/SGM Summer Project/2011/06/28

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Check overnight growth (7x200ml)

Part Growth Colonies
BBa_I13453 Yes 3
BBa_I15010 No NA
BBa_B0034 Yes 14
BBa_I15008 No NA
BBa_I15009 Yes 1
BBa_R0040 Yes 3
BBa_B0015 yes 2

Inoculation

Only 5 parts were used to incoulate, BBa_I13453, BBa_B0034, BBa_I15009, BBa_R0040 and BBa_B0015

    1. 2 10ml tubes for each (10 total)
    2. Label the tubes
    3. Turn on the flame
    4. I13453, add 10μL Ampicillin
    5. B0034, add 10μL Ampicillin
    6. I15009, add 50μL Kanamycin
    7. R0040, add 10μL Ampicillin
    8. B0015, add 10μL of Ampicillin and 50μL of Kanamycin
    9. Inoculate the 5 plates into each 2x 10 ml tubes.
    10. 10 tubes are then left in the shaker overnight

Transformation of BBa_I15008 and I15010

The plasmids are transformed with 2 different competent cells, XL-1-Blue and DH5-α.

  1. Fill up the box with ice.
  2. Take 4 eppendorf tubes, 2x XL-1-Blue and 2x DH5-α from the -80°C freezer and put it on ice.
  3. Take the plasmids (Igem plates 2010. If the plates are not used, meaning the well of interest is not puncrures, add 10μL of free nuclease water)from the -20°C freezer
  4. Take out the Soc from the -20°C freezer°C and leave in the incubater for 10 min
  5. Add 2μL, plasmid and transfer into each 4 eppendorf. Mix the DNA to the cells by vortex and leave in the ice for 5 min. Note= while doing this, flame should be on.
  6. Take 4 eppendorf tubes into 42°C waterbath and leave it for 1 min exactly
  7. Add 250μL of Soc.
  8. The tubes are then left in the shaker for 1 hr
  9. Take 4 selective agar plates and leave it in the incubater upside down with lid open slightly to lit air through.
  10. Take out the tubes from the shaker
  11. Take out the plates from the incubater.
  12. Turn on the flame. Transfer the contents from each epppendorf tubes into each 4 plates.
  13. Add glass beads.
  14. Now take all 4 plates and shake for 8-20 sec
  15. Remove the glass beads
  16. The plates are then left overnight in incubater at 37°C




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