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	###300uL C0060		###300uL C0060
	##Place into 37deg C incubator at 3:55PM		##Place into 37deg C incubator at 3:55PM
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		+	Next: miniprep and screen the DNA.

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Revision as of 12:50, 6 September 2007

McGill Home

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Contents 1 May 2007 1.1 May 14 1.2 May 15 1.3 May 16 1.4 May 30 2 June 2007 2.1 June 4 2.2 June 20

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.

1. 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)

1. Transformation of extracted DNA
   1. Chill cells on ice for 10 min
   2. Heat shock cells at 42deg C for 30 sec
   3. Add DNA for cells
   4. Add 500uL of 42deg C SOC medium to each vial
   5. Incubate on ice for 1 min
   6. Place in 37deg 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 37deg C incubator at 3:55PM

Next: miniprep and screen the DNA.