Standard Protocols
From 2006.igem.org
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=== Transformation into Competent Cells === | === Transformation into Competent Cells === | ||
Revision as of 15:23, 5 July 2006
Resuspension of Dry DNA
- Puncture a hole through the foil with a pipette tip
- Add 15 ul of distilled water
Transformation into Competent Cells
- Take 1ul of suspended DNA and add to an aliquot of competent E-Coli cells (taken from the freezer at -80°C)
- Mix gently and leave in ice bucket for 10 to 30 minutes
- Place aliquot in heat block set to a temperature of 42°C for approximately 2 minutes (no longer than 3 minutes)
- Return to ice for a short period of time (approximately 3 minutes)
- Add 1 ml of LB liquid media
- Shake for about an hour at 37°C
- Plate on agar containing the correct antibiotic
- Incubate at 37°C overnight
- Remove from oven and rest at room temperature for approximately an hour
- Move to refrigerator
