Standard Protocols
From 2006.igem.org
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# Add 1 ml of LB liquid media | # Add 1 ml of LB liquid media | ||
# Shake for about an hour at 37°C | # Shake for about an hour at 37°C | ||
| - | # Plate on agar containing the correct antibiotic | + | # 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 |
# Incubate at 37°C overnight | # Incubate at 37°C overnight | ||
# Remove from incubator and rest at room temperature for approximately an hour | # Remove from incubator and rest at room temperature for approximately an hour | ||
Revision as of 13:59, 6 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, put approx. 100ul of cells on one plate, and the remainder on the other after centrifuging and resuspending the pellet
- Incubate at 37°C overnight
- Remove from incubator and rest at room temperature for approximately an hour
- Move to refrigerator
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