Electrophoresis and Restriction Enzymes

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Restriction Enzymes and Electrophoresis

Bhumik Patel


Restriction enzymes are tools in DNA research that can cut DNA into exactly needed pieces. Certain cuts can be rough, while others can be clean. Certain cuts can have an organized pattern to have a staggered cut. Other cuts will leave complementary bases with them. Electrophoresis allows the manipulation of DNA to separate and organize those parts. Electrophoresis is the substrate electric movement of the separation of DNA. Gel Electrophoresis can separate both DNA and RNA. This can also allow the knowledge of the unknown bases of a strand of DNA. This can allow the knowledge of familiar patterns that can create a unique signature. Separation of these molecules depends on the charge of the molecule and the mass. The charges are shown toward the opposite charge and it will attract to. The gel that organizes the DNA fragments is agarose and polyacrylamide. Polyacrylamide is a material that is similar that is found in soft contact lenses. DNA fingerprinting is a way of showing differences in DNA from one being to another. 99% of the human DNA is similar to all other humans. The remaining 1% is enough to tell us the differences from one to another. The one-percent has a very large possibility of options for your DNA base sequence. We were to determine that electrophoresis can show differences in DNA.

We had used in this lab 0.8% agarose gel, gel tray, TBE Running Buffer 1X (350ml), DNA Strain, Staining Tray, Micropipets, Metric Ruler, semi-log graph paper, goggles, aprons, and gloves. We shared all the DNA samples.

Molding the gel:
1) Add rubber stoppers to the ends of the gel plate
2) Add well placements
3) Add 25ml of agarose gel to gel plate
4) Add gel stain
5) Let the gel rest for about 20 minutes or until opaque.

Loading and running the gel:
1) Remove rubber stoppers and well placement from the gel plate. (remove gel placement straight up) 2) Add...
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