Gel Electrophoresis. Definition – COPY ME! Separation of DNA fragments according to size and charge Based on movement through a gel medium when an.

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Presentation transcript:

Gel Electrophoresis

Definition – COPY ME! Separation of DNA fragments according to size and charge Based on movement through a gel medium when an electric field is applied.

DNA negatively charged Organic molecules such as DNA are charged. DNA is negatively charged because the phosphates (red circles) that form the sugar-phosphate backbone of a DNA molecule have a negative charge

DNA cut up using restriction endonucleases

Make up the gel which the DNA will be put into

A comb is put in the gel to create holes which we call wells

Dye added to the DNA Makes the sample visible when it is put into the agarose wells

Buffer solution added to the tank This ensures that the electric current goes through the whole tank and that maintains that ions can move in the solution

DNA samples loaded into wells

Electrical current applied to the chamber Safety cover is put over the top and the current is switched on The dye will migrate through the gel toward the positive electrode, as will the DNA Depending on how much voltage is applied and how warm the gel is and size and shape of molecules will depend on how fast the molecules move through the gel Smaller fragments will move easier so they will be closer to the positive electrode Once the dye has moved through the gel to the buffer, the electrical current is switched off and gel is removed from the tray

DNA is stained using ethidium bromide Gel is stained using ethidium bromide which binds to DNA it shows up as bands in UV light Small molecules are at the bottom of the gel and large ones stay nearest to the wells

Let’s act it out!

Group Activity!

Now try your Gel Electrophoresis sheet! (Biozone 1 –p252)