IGEM:Harvard/2010/General Protocols/DNApurification

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(New page: *****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 flas...)
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*Load samples into the gel with a 100 kb+ ladder
*Load samples into the gel with a 100 kb+ ladder
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[[image:Onekbplusladder.jpeg]]
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*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
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*Image the gel, cut desired parts out of the gel with a scalpel, and proceed to the Qiagen DNA purification kit.

Revision as of 11:09, 7 June 2010

          • 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

image:Onekbplusladder.jpeg

  • 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.
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