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)
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, put approx. 100ul of cells on one plate, and the remainder on the other after centrifuging and resuspending the pellet
8. Incubate at 37°C overnight
9. Remove from incubator and rest at room temperature for approximately an hour
10. Move to refrigerator

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