Standard Protocols

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Revision as of 15:22, 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