User:Jose Fabricio Lopez/Notebook/GEDF/Protocols

This page contains all protocols and basic techniques to work on Trichoderma spp.

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.