Standard Protocols

From 2006.igem.org

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)

and transform into your favorite competent cells, plate out on a plate with the correct antibiotic and grow overnight. Your goal here is to obtain single colonies.

      4. Pick a single colony and inoculate some broth (with the correct antibiotic) and grow ~18 hours.
      5. Use the resulting culture to miniprep AND make your own glycerol stock (for further instruction on making a glycerol see this page).