Team:UNIPV-Pavia/Protocols/Precipitation

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

DNA precipitation with sodium acetate (estimated time: 1 hour) Materials needed:


 * sodium acetate
 * absolute ethanol
 * ethanol 70%
 * ddH2O
 * previously cut plasmid
 * Add 1/10 digestion volume of sodium acetate 3 M
 * Add 2.5 digestion volume of absolute ethanol
 * Freeze at -80°C for 30 min
 * Centrifuge at 13000 rpm, 4°C for 20 min
 * Decant supernatant
 * Add 50 µl of 70% ethanol
 * Centrifuge at 13000 rpm, 4°C for 20 min
 * Remove all supernatant with a pipette
 * Air dry pellet until ethanol is totally removed
 * Elute with 5 µl ddH2O