Difference between revisions of "Agarose Gel Electrophoresis"
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== Purpose == | == Purpose == | ||
| + | This protocol is used to purify strands of DNA | ||
== Materials Needed == | == Materials Needed == | ||
| + | -Electrophoresis gel | ||
| + | |||
| + | -Electrophoresis tank | ||
| + | |||
| + | -Power supply | ||
| + | |||
| + | -Buffer | ||
| + | |||
| + | -Loading Dye | ||
| + | |||
| + | -1 kb marker | ||
| + | |||
| + | -Micropipette | ||
| + | |||
| + | -UV light source | ||
== Protocol == | == Protocol == | ||
| + | -Place the hardened gel into the electrophoresis tank with the wells closest to the negative end | ||
| + | |||
| + | -Add enough buffer to cover the gel until the wells are covered | ||
| + | |||
| + | -Load 10 microliters of a 1 kb sample into the first well as a marker using a micropipette | ||
| + | |||
| + | -Mix 10 microliters of the DNA you want to run with .1 microliters of loading dye | ||
| + | |||
| + | -Load the mixture into the second well using a micropipette | ||
| + | |||
| + | -Repeat the above two steps for as many samples as you have | ||
| + | |||
| + | -Attach the nodes from the power supply to the electrophoresis tank | ||
| + | |||
| + | -Set the voltage to the desired level (1-10 V/cm of gel) | ||
| + | |||
| + | -Turn on the power | ||
| + | |||
| + | -When the blue dye from the loading buffer has travelled far enough to allow separation of the DNA fragments, turn off the power | ||
| + | |||
| + | -Use a UV light source to observe the bands | ||
== Notes == | == Notes == | ||
Latest revision as of 12:52, 13 July 2015
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Contents
[hide]Purpose
This protocol is used to purify strands of DNA
Materials Needed
-Electrophoresis gel
-Electrophoresis tank
-Power supply
-Buffer
-Loading Dye
-1 kb marker
-Micropipette
-UV light source
Protocol
-Place the hardened gel into the electrophoresis tank with the wells closest to the negative end
-Add enough buffer to cover the gel until the wells are covered
-Load 10 microliters of a 1 kb sample into the first well as a marker using a micropipette
-Mix 10 microliters of the DNA you want to run with .1 microliters of loading dye
-Load the mixture into the second well using a micropipette
-Repeat the above two steps for as many samples as you have
-Attach the nodes from the power supply to the electrophoresis tank
-Set the voltage to the desired level (1-10 V/cm of gel)
-Turn on the power
-When the blue dye from the loading buffer has travelled far enough to allow separation of the DNA fragments, turn off the power
-Use a UV light source to observe the bands
Notes
Papers where this or a similar method has been used
<PMIDsummary />
Related Ontology Terms
References
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