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After having done a sequencing reaction, it is necessary to run a gel
to see and read the sequence. In order to read the maximum sequence (400
bases), we usually do a DOUBLE RUN. The four reactions are loaded and run
for a length of time (usually 1:30 h) and then they are reloaded in another
set of wells, and the gel can run another 2 or more hours, depending on
the wattage.
When it comes to reading the gel, one will start with the second run, which will show the beginning of the sequence (hopefully), and finish with the first run which will show the end of the sequence. |
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Toward the top of the gel, it is often very difficult to interpret the results, because the bases are very close together. A good double run will show a minimum of overlapping sequence.