Team:Calgary/7 July 2010


Revision as of 19:58, 7 July 2010 by Jdchoo (Talk | contribs)

Wednesday July 7, 2010


  • Today I completed the transformation of my construct (R0040-I0504) and plated them in 6 different Ampicilin Resistant plates.

Procedure: Transformation

  • 1. Thaw Competent Cells
  • 2. Add 20 uL DNA
  • 3. Ice for 1/2 hour, Heat Shock 5 minutes at 37 degrees Celcius, Ice 5 min.
  • 4. Recover with 250 uL SOC for 30 minutes
  • 5. Spin down at 14 RPM, Concentrate to approximately 100 uL
  • 6. Plate 25 uL on plate 1,2,3 and Plate 50 uL on plate 4,5,6
  • 7. Leave in Incubator overnight for 20 hours

The plates are labeled as following:

  • P1: R0040-I13504 (construct tube C 6 July) 25 uL
  • P2: R0040-I13504 (construct tube D 6 July) 25 uL
  • P3: R0040-I13504 (construct tube F 6 July) 25 uL
  • P4: R0040-I13504 (construct tube C 6 July) 50 uL
  • P5: R0040-I13504 (construct tube D 6 July) 50 uL
  • P6: R0040-I13504 (construct tube F 6 July) 50 uL

Then I left the plates in the Incubator which is at 37 degrees Celcius. I will leave it there until 10:00 am tomorrow (July 8th), which would let the cells grow for 20 hours.

Jeremy: Today I completed the plasmid switch for I0500 into pSB1AK3, mini prep I0500+B0034 with Sigma-Aldrich and finished a restriction digest of EcoRI and PstI on I0500+B0034

  • At the end of the heat killing process, I added

5 uL Antartic Phosphotase Buffer 5 uL of Water and 1 uL Antartic Phosphotase It was placed in the water bath at 37°C for half an hour and then I ligated the insert and the vector using 5 uL of Insert and 5 uL of Vector 1 uL Quick Ligase 10 uL 2x Quick Ligase Buffer The construct is left in the fridge overnight and transformation will be done tomorrow.

No notebook page exists for this date. Sorry!