What is Gel Electrophoresis?
- Electrophoresis- A method for separating large molecules by placing them in a gel in the presence of an electric field
- Used to separate DNA, RNA, and Proteins
- Extraction – DNA is removed from white blood cells, hair root cells, or tissue
- Restriction – DNA is cut with restriction enzymes into fragments
- Separation – DNA is negatively charged and is naturally attracted to the positive side of the gel, repelled by the negative ends of the gel. DNA fragments travel through micro-pores in the gel.
- Separation is based on 3 characteristics- Charge, size, and shape.
- Mix agarose, buffer, and water
- Buffer – Controls pH, Conducts electricity
- Microwave for 1 minute, swirl
- Repeat step 2 until solution is clear
- Place flask in water bath to lower temp to 55 degrees Celsius
- Pour solution into molds until level with rubber stoppers
- Holds exactly 40 microliters
- DO NOT TURN UPSIDE DOWN!
- To fill Micropipette – Depress, Hold, Insert, Release, Remove
- To fill Gel – Insert, Depress, Hold, Remove, Release