"

 

From 2009.igem.org

Home The Project Motivation Solution Results & Conclusions References Materials & Methods Measurements Instruments Protocols Software Developed Parts Submitted Characterization The Team Notebook Week-book Freezer Management Biological Safety Pavia History University Gallery Sponsors Gallery Team Locations		Protocols
	LB medium DNA resuspension from iGEM 2009 plates Transformation X-Gal staining protocol for beta galactosidase (blue/white screening) Glycerol stocks Plasmid extraction BioBrick digestion with restriction enzymes Electrophoresis DNA gel extraction DNA precipitation with sodium acetate Ligation PCR Sensing ethanol concentration: our do-it-yourself kit

Electrophoresis

(estimated time: 2 hours)

Prepare agarose gel in 1x TBE buffer Add ethidium bromide (using gloves and face mask for your safety): 1 µl in the small size agarose gel (70 ml) 2 µl in the middle size agarose gel (150 ml) 4 µl in the big size agarose gel (250 ml) Cast the gel, insert the well-forming comb and let it polymerize Add the loading buffer to each sample Load the samples and 8 µl of marker Set to 70-100 volts and electrophorese for the required amount of time Use UV-light to look at the bands Take a picture of the gel, if needed (not when bands have to be cut!!!)

DNA samples Ethidium bromide Loading buffer 10x Blue Juice, Invitrogen TBE (Tris/Borate/EDTA buffer) 5x (final volume 1 liter) 54 gr Tris 27.5 gr Borate 20 ml EDTA 0.5 M (pH 8) Marker (1 kb DNA Ladder, Promega) Face mask and gloves Electrophoresis apparatus Transilluminator

• NOTE: when not specified, the marker has the following pattern:

• Recent changes
• What links here
• Related changes
• Special pages
• My preferences

• Permanent link
• Privacy policy
• Disclaimers