1 (A) Agarose Gel Electrophoresis

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Gel electrophoresis

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technique used to seperate individual components of a mixture of molecules using an electrical current

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In gel electrophoresis molecules must be ...

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water soluable and have at least a slight electrical charge

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agarose

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gelatin-like material

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buffer

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electrically conductive solution

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the buffer stabalizes the pH of the molecules to be seperated and controls the amount of electrical current that passes through the chamber

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when an electrical current is passed through the chamber...

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the molecules in the mixture will begin to move towards the positive or negative poles of the chamber.

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Molecules with a negative charge will migrate toward posotive pole and those with a positive charge will migrate towards negative pole.

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The agarose gel slows the movement of the molecules and acts like a sieve

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slows the movement
sieve

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Outline the process of Gel Electrophoresis

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Extracting the DNA
Separating the DNA
Visualising the DNA
Isolating the gene of interest

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Extracting the DNA

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DNA extracted

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Separating the DNA

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DNA cut by restriction enzymes placed in wells in polyacrylamide gel in buffer solution with electrodes at either end

DNA fragments travel along the gel to the ANODE or positively charged end according to size. DNA fragments thus separated

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Visualising the DNA

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Polyacrylamide gel contains ethidium bromide which unites with the DNA and fluoresces under UV Light
DNA placed in dark room and UV light shone on DNA so can see fragments

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Isolating the gene of interest

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A marker is loaded onto the gel
Details size of gene of interest
Marker ocmpared to fragments to find gene of interest

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Fate of the DNA after gel elecrtophoresis?

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The gene of interest could be removed from the gel, purified and then cloned