Penn State University 2006:Transformation
From 2006.igem.org
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='''Transformation'''= | ='''Transformation'''= | ||
''Time: 2.25 hr'' | ''Time: 2.25 hr'' | ||
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===<font color="blue">Media/Reagents</font color>=== | ===<font color="blue">Media/Reagents</font color>=== | ||
Latest revision as of 00:33, 24 June 2006
Contents |
Transformation
Time: 2.25 hr
Media/Reagents
- Competent cells
- SOC1
- Antibiotic LB plates
Pre
- Chill ligated DNA on ice
- Thaw competent cells ((found in eppendorfs in box on second-shelf down of -80ºC freezer) on ice for ~5 min.
Transformation
- Add 3 μL ligation mixture to cells in bottom of tube
- Incubate on ice for 30 min
- Heat shock in 42ºC water bath for 1 min (warm SOC media at same time).
- Incubate on ice for 2 min
- Add 500 μL warm SOC media
- Shake in 37ºC room for 1 hr
- Spread 500 μL on LB/antibiotic plates using rotator and glass spreading wand.
- Incubate plates in 37ºC room overnight.
1To make SOC, add 200 μL 500 mM filter-sterilized glucose to 5 mL SOB (20 mM final glucose concentration)
