Gel Electrophoresis
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How to cut DNA:
Restriction enzymes are used to "cut" DNA at specific points. Once the enzyme binds to the recognition sequence (e.g. between C and G), restriction enzyme cuts the sugar-phosphate groups of the DNA strands.
Restriction enzymes are used to "cut" DNA at specific points. Once the enzyme binds to the recognition sequence (e.g. between C and G), restriction enzyme cuts the sugar-phosphate groups of the DNA strands.
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How to make the gel for electrophoresis:
Step 1: Put a small amount of agarose into a flask.
Step 1: Put a small amount of agarose into a flask.
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Step 2: Then add some liquid buffer to the flask to enable the electrical charges to run through the gel. Heat the mixture until the agarose blends in with the liquid buffer.
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Pour the mixture into a mold. Put the comb into the gel liquid and let it cool down and tiny holes will form inside the gel.
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How to set up the apparatus of gel electrophoresis:
Pour some liquid buffer into an electrophoresis box and put the mold into the box.
Pour some liquid buffer into an electrophoresis box and put the mold into the box.
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Use a micropipette to put uploading buffer into the DNA sample so that the DNA become dyed thicker. It's easier to see.
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Suck up the DNA sample (with loading buffer) with a micropipette and eject it into the wells of the gel.
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How to run the gel?
Connect the black cord from the electrophoresis box to the negative end of the power supply and the red cord from the box to the positive end and let an electric current run through. Notice while the process is taking place, there are bubbles that come out of the electrodes.
Connect the black cord from the electrophoresis box to the negative end of the power supply and the red cord from the box to the positive end and let an electric current run through. Notice while the process is taking place, there are bubbles that come out of the electrodes.
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How to stain the gel and analyse the results:
DNA staining solution is a chemical called ethidium bromide which binds to DNA and shows up under fluorescent light.
DNA staining solution is a chemical called ethidium bromide which binds to DNA and shows up under fluorescent light.
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Put the gel into the staining solution. After approximately half an hour, remove the gel and place it onto the UV light box.
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DNA strands are measured in base pairs (bp) as in how many base pairs there are in a cut DNA strand.
Source: http://learn.genetics.utah.edu/content/labs/gel/