Team:UNIPV-Pavia/Protocols/Resuspension

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

Plasmid resuspension from IGEM paper spots (estimated time: 25 min + 5 min for every part if you use scalpel/tweezers or + 15 min for every part if you use punch tool) Materials needed:
 * Pre-warmed at 42°C TE
 * Desired spot location information
 * Scalpel and tweezers (or punch tool)
 * ddH20
 * 99% ethanol
 * 0.5 ml tubes
 * Put 10 µl of pre-warmed TE into a 0.5 ml tube.
 * Cut paper spots using scalpel and tweezers (or punch tool, following the instructions provided with the IGEM kit); if you use scalpel and tweezers, try to cut pieces of about the same dimension of the punch tool.
 * Put the cut paper into the 0.5 ml tube.
 * Clean scalpel and tweezers (or punch tool) with water and ethanol every time you cut a spot; be careful to dry your tools correctly, especially if you use punch tool, which needs much more time to dry than scalpel/tweezers.
 * Incubate at 42°C for 20 min.
 * Vortex and spin down.