![]() Loading too much DNA will make it difficult to obtain crisp bands and analyze the results. If your digest lanes look like your uncut lane then there is something wrong! This is due to the fact that the circular DNA takes on several conformations the most abundant being: supercoiled, relaxed and nicked. When uncut plasmid DNA is isolated and run on an agarose gel, you are likely to see 3 bands. Always run control uncut DNA to ensure your enzymes are working.Choose your ladder based on the expected band sizes. Ladders allow you to interpret the bands that you get in your sample lanes. ![]() Bonus: The dyes also run at predicted sizes so you can estimate how far down the gel your bands have traveled based on the dye! The glycerol in the buffer will make sure your sample settles in the gel well and the dyes provide a visual reference point so you can easily assess how far the gel has run.
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