"
Team:Calgary/Notebook/Protocols/BacterialTransformation
From 2013.igem.org
(Difference between revisions)
Wkeithvan (Talk | contribs)
(Created page with "<html> <div id="Banner"><h1>Bacterial Transformation</h1></div> </html> {{Team:Calgary/ContentPage}} <html> <section id="Content"> <h1>Bacterial Transformation</h1> <h2>Reagents...")
Newer edit →
(Created page with "<html> <div id="Banner"><h1>Bacterial Transformation</h1></div> </html> {{Team:Calgary/ContentPage}} <html> <section id="Content"> <h1>Bacterial Transformation</h1> <h2>Reagents...")
Newer edit →
Revision as of 06:52, 24 September 2013
Bacterial Transformation
Bacterial Transformation
Reagents and Materials
- Chemically competent E. coli
- DNA (e.g., ligation reaction)
- SOC medium
- LB agar plates + desired antibiotic
Protocol
- Thaw 100μL of competent cells (per transformation) on ice just before they are needed
- Add DNA (max 20μl) thawed cells and mix by flicking the side of the tube. Leave on ice for 30 minutes
- Heat shock 5 minutes at 37 degrees Celsius
- Place on ice for 5 minutes
- Add 250μl SOC medium to each tube
- Incubate for 30 to 60 minutes with shaking at 37 degrees Celsius (note that for Kanamycin containing plasmids always use one hour)
- Spin down to remove all supernatant except approximately 100 μL
- Plate approximately 50 μL on antibiotic plates
- Grow overnight at 37 degrees Celsius
