IGEM:Harvard/2010/General Protocols/DNApurification: Difference between revisions
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*Load samples into the gel with a 100 kb+ ladder | *Load samples into the gel with a 100 kb+ ladder | ||
[[image: | [[image:1kbLadderigem2010.JPG]] | ||
*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 | *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. | *Image the gel, cut desired parts out of the gel with a scalpel, and proceed to the Qiagen DNA purification kit. | ||
Latest revision as of 07:03, 17 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
- 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.
