Standard Protocols
From 2006.igem.org
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=== Transformation into Competent Cells === | === 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 | + | # 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 | ||
Revision as of 15:14, 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
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).
