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Revision as of 12:52, 6 September 2007 (view source) Tim J. (Talk | contribs) (→June 20) ← Older edit		Revision as of 12:56, 6 September 2007 (view source) Tim J. (Talk | contribs) Newer edit →
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	=== May 30 ===		=== May 30 ===
	# Test PCR Machine		# Test PCR Machine
		+
	== June 2007 ==		== June 2007 ==
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	## 200mL of 2x LB		## 200mL of 2x LB
	## 2mL of Kanamycin		## 2mL of Kanamycin
-
	=== June 20 ===		=== June 20 ===
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	II - Transformation of extracted DNA		II - Transformation of extracted DNA
	#Chill cells on ice for 10 min		#Chill cells on ice for 10 min
-	#Heat shock cells at 42deg C for 30 sec	+	#Heat shock cells at 42°C for 30 sec
	#Add DNA for cells		#Add DNA for cells
-	#Add 500uL of 42deg C SOC medium to each vial	+	#Add 500uL of 42°C SOC medium to each vial
	#Incubate on ice for 1 min		#Incubate on ice for 1 min
-	#Place in 37deg C shaker incubator for 1h 30min	+	#Place in 37°C shaker incubator for 1h 30min
	#Plate onto 4 separate amp plates		#Plate onto 4 separate amp plates
	##25uL R0062		##25uL R0062
Line 198:		Line 198:
	##400uL R0062		##400uL R0062
	##300uL C0060		##300uL C0060
-	#Place into 37deg C incubator at 3:55PM	+	#Place into 37°C incubator at 3:55PM
	'''Next''': miniprep and screen the DNA.		'''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 ===

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Revision as of 12:56, 6 September 2007

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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 2.3 June 21 2.4 June 22

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