Team:UNIPV-Pavia/Protocols/AntarcticPhosphatase

The protocols we used


 * LB medium preparation
 * Plasmid resuspension from IGEM paper spots
 * Transformation
 * Plasmid extraction
 * BioBrick digestion with restriction enzymes
 * DNA gel extraction
 * DNA precipitation with sodium acetate
 * Antarctic Phosphatase
 * Ligation
 * PCR

Antarctic Phosphatase (estimated time: 1 hour and 30 min) Materials needed:


 * NEB Antarctic Phosphatase
 * 10X NEB Antarctic Phosphatase buffer
 * Cut and gel-extracted vector
 * Add the proper amount of 10X buffer to a final concentration of 1X (e.g. 2 µl of 10X buffer in a final volume of 20 µl).
 * Add 1 µl of Antarctic Phosphatase (up to 5 µg of cut vector).
 * Incubate at 37°C for 1 hour (Antarctic Phosphatase works).
 * Incubate at 65°C for 15 min (Antarctic Phosphatase inactivation).