IGEM:Harvard/2010/General Protocols/DNApurification

• • • • • NOTE: Ethidium Bromide (EtBr) is a mutagen ---- always wear gloves when handling it*****
• Making an agarose gel
  • To make a 1% gel, add 1g agarose to 100mL 1x TAE buffer in 200mL flask
  • Microwave for 1.5 minutes
  • Swirl and check that the agarose is completely dissolved and there are no air bubbles
  • Allow gel solution to cool for a bit
  • While gel is cooling, tape the ends of gel mold so gel liquid cannot escape. Don't forget the combs!! Sit mold on a flat surface.
  • Add 4.5ul of EtBr to 100mL of gel and mix by swirling (avoid making bubbles)
  • Pour into taped gel mold and allow to cool until solid

• Load samples into the gel with a 100 kb+ ladder

• Place the gel in the electrophoresis chamber and run the gel at 100V for 30 minutes until the DNA loading dye has run partially through the gel
• Image the gel, cut desired parts out of the gel with a scalpel, and proceed to the Qiagen DNA purification kit.