Difference between revisions of "DNA sequencing"
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To make a gel, add agarose to room temperature 1x TAE buffer, then heat in the microwave to dissolve it into solution. It is important to make it completely clear - no grains of agarose should be visible when you swirl the
To use the gel, put it into a
Optionally, you can use one of your lanes for a
Run the gel at 100 volts DC for 20 minutes. If you leave it for 30 minutes or longer, you will lose your results. Higher voltages work more quickly but give bands that are less sharp.
Visualize the gel using a
If you get a strong or medium-strong clear band, it is very likely that you will get a clean sequence from the sample. If you get a weak band, smear or no band at all,
TAE buffer is the standard buffer used, but it may not be the best choice - sodium borate can be used instead, allowing higher voltage and therefore faster gels. See