Standard Protocols

From 2006.igem.org

(Difference between revisions)
Jump to: navigation, search
(Resuspension of dry DNA)
Line 3: Line 3:
# Puncture a hole through the foil with a pipette tip
# Puncture a hole through the foil with a pipette tip
# Add 15 ul of distilled water
# Add 15 ul of distilled water
 +
=== Transformation into Competent Cells ===
=== Transformation into Competent Cells ===

Revision as of 15:23, 5 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 oven and rest at room temperature for approximately an hour
  10. Move to refrigerator
Personal tools
Past/present/future years