McGill/Team 1: Fluorescence Complementation

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May 2007
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June 2007
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Contents

May 2007

May 14

  1. Made the same as May 15, but did not use Type 1 (milliQ) H2O, so the solutions were discarded.

May 15

  1. Made 1M CaCl2, and glycerol/CaCl2 solutions
    1. 1M CaCl2 Solution: 100mL was made
      1. CaCl2 Dihydrate = 146.986 g/mol
      2. 100mL of 1M CaCl2 requires 14.699g CaCl2 Dihydrate
    2. Glycerol/CaCl2 Solution: 30mL was made
      1. 5.00mL of 60% glycerol solution
      2. 3.00mL of 1M CaCl2
      3. 22.00mL of Type 1 (milliQ) H2O

May 16

  1. Transformation of Cells:
    1. Transformed 1 vial of cells with Jun vector, and another vial with Fos vector (1uL of vector each); Sterile
    2. Used 400uL of LB (sterile) for each vial
    3. Incubated for 1 hour in shaker at 37C, then incubate overnight at 37C

May 30

  1. Test PCR Machine


June 2007

June 4

  1. Made Kanamycin plates
    1. 200mL of Agar
    2. 200mL of 2x LB
    3. 2mL of Kanamycin

June 20

Today: extract DNA from plates, transform, plate on amp plates.

I - Extraction of DNA from Plate (5M and 9G)

  1. Puncture foil with pippette tip
  2. Add 15uL of Type 1 water
  3. Remove all liquid (with DNA in solution)

II - Transformation of extracted DNA

  1. Chill cells on ice for 10 min
  2. Heat shock cells at 42°C for 30 sec
  3. Add DNA for cells
  4. Add 500uL of 42°C SOC medium to each vial
  5. Incubate on ice for 1 min
  6. Place in 37°C shaker incubator for 1h 30min
  7. Plate onto 4 separate amp plates
    1. 25uL R0062
    2. 25uL C0060
    3. 400uL R0062
    4. 300uL C0060
  8. Place into 37°C incubator at 3:55PM

Next: miniprep and screen the DNA.

June 21

Checked colonies in the 37°C incubator. Well formed colonies were on each plate, so plates (all 4) were transferred to the small fridge (middle shelf on the right).

June 22