"

 

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)

Materials needed:

• DNA samples
• Ethidium bromide
• Loading buffer 10x Blue Juice, Invitrogen
• TBE (Tris/Borate/EDTA buffer)
  • TBE 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

• Prepare 1% 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 (220 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 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)

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

• Printable version
• Permanent link
• Privacy policy
• Disclaimers