User:Jose Fabricio Lopez/Notebook/GEDF/Protocols: Difference between revisions
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1.-Introduce cells in DNA solution. | 1.-Introduce cells in DNA solution. | ||
2.-Put cells in ice. | 2.-Put cells from REVCO in ice and Cuvettes too. | ||
3.-Select Cuvettes for electroporation and put voltage. Wait until sound occurs. | 3.-Select Cuvettes for electroporation and put voltage according with the cuvette. Wait until sound occurs. | ||
4.-Put 500μL of LB liquid (without antibiotic). | 4.-Put 500μL of LB liquid (without antibiotic). | ||
Latest revision as of 15:24, 10 January 2012
Protocols
DNA
Transformation for E. Coli by electrophoration
Reagent and instruments
- LB medium Liquid
- LB medium Solid
- DNA of plasmid. (10μL)
- Electrocompetent cells.
- Ice.
- Deionized water.
- Paper for deinization.
- Antibiotic
Instrucctions
1.-Introduce cells in DNA solution.
2.-Put cells from REVCO in ice and Cuvettes too.
3.-Select Cuvettes for electroporation and put voltage according with the cuvette. Wait until sound occurs.
4.-Put 500μL of LB liquid (without antibiotic).
5.-Grow up cells by 1 hour at 37°C mixing.
6.-Petri dishes with antibiotic.
7.-Plate cells.
Remember: Use controls for medium and cells.
