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Process of Electrophoresis Extracted from http://www.bio-rad.com/webroot/web/pdf/lse/literature/1660077EDU.pdf Video preview Using a micropipette http://www.cleanvideosearch.com/media/action/yt/watch?videoId=p-OPOYbeZP0 Agarose gel electrophoresis http://www.cleanvideosearch.com/media/action/yt/watch?videoId=vq759wKCCUQ DNA electrophoresis sample loading http://www.cleanvideosearch.com/media/action/yt/watch?videoId=tTj8p05jAFM Agarose gel electrophoresis is a procedure used to separate DNA fragments based on their sizes. DNA is an acid and has many negative electrical charges. Scientists have used this fact to design a method that can be used to separate pieces of DNA. A solution containing a mixture of DNA fragments of variable sizes is placed into a small well formed in an agarose gel that has a texture similar to gelatin. An electric current causes the negatively-charged DNA molecules to move towards the positive electrode. Imagine the gel as a strainer with tiny pores that allow small particles to move through it very quickly. The larger the size of the particles, however, the slower they are strained through the gel. After a period of exposure to the electrical current, the DNA fragments will sort themselves out by size. Fragments that are the same size will tend to move together through the gel and form bands. This lab can also be done using “model” DNA (AKA dye). The key difference is that dye molecules are very similar in size, but they differ in the amount of electrical charges. Black Red DNA Gel Box Filled with buffer solution to conduct electricity. To conduct current effectively through the gel, the box must be filled with enough buffer so that the gel is under 1-2mm depth of buffer. Gels using “quick run buffer” can be run at 170V. Teacher notes below Teacher notes Yes can use just food coloring, but it does tend to trail during loading (might set max value at 18microliters Food coloring Dye concentration suggestions Darker colors 10-5% food coloring 90-95% water (V/V) Lighter food coloring 15-10% food coloring 85-90% water (v/v) Use 0.25X TAE (quick run buffer) --to prep 0.25X TAE from 50X TAE stock solution Either from 10ml of stock TAE +1990ml of dH2O or, 1ml of stock TAE +199ml of dH2O -set up gel boxes mostly full w/ TAE. Each group gets a small bottle of TAE to add that last little bit of TAE so 1-2mm above -probably change TAE solution is reusable but will probably need changing every 2-3 labs