Standard Protocols

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(Transformation into Competent Cells)
Line 15: Line 15:
# Plate on agar containing the correct antibiotic
# Plate on agar containing the correct antibiotic
# Incubate at 37°C overnight
# Incubate at 37°C overnight
-
# Remove from oven and rest at room temperature for approximately an hour
+
# Remove from incubator and rest at room temperature for approximately an hour
# Move to refrigerator
# Move to refrigerator
[http://2006.igem.org/University_of_Edinburgh_2006 Main page]
[http://2006.igem.org/University_of_Edinburgh_2006 Main page]

Revision as of 12:44, 6 July 2006

Resuspension of Dry DNA

  1. Puncture a hole through the foil with a pipette tip
  2. Add 15 ul of distilled water


Transformation into Competent Cells

  1. Take 1ul of suspended DNA and add to an aliquot of competent E-Coli cells (taken from the freezer at -80°C)
  2. Mix gently and leave in ice bucket for 10 to 30 minutes
  3. Place aliquot in heat block set to a temperature of 42°C for approximately 2 minutes (no longer than 3 minutes)
  4. Return to ice for a short period of time (approximately 3 minutes)
  5. Add 1 ml of LB liquid media
  6. Shake for about an hour at 37°C
  7. Plate on agar containing the correct antibiotic
  8. Incubate at 37°C overnight
  9. Remove from incubator and rest at room temperature for approximately an hour
  10. Move to refrigerator


[http://2006.igem.org/University_of_Edinburgh_2006 Main page]

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