![]() TBE buffer has a higher buffering capacity than TAE buffer. Higher molecular weight DNA separates better with a lower percentage gel. Avoid overfilling gel wells to prevent smearing of DNA down the surface of the gel. If you see DNA migration issues or smearing after post-staining with GelRed® or GelGreen®, then the problem is not caused by the nucleic acid dye.Other tips to improve agarose gel resolution: See the GelRed® Product Information Sheet or GelGreen® Product Information Sheet for detailed protocols. While we recommend post-staining gels for 30 minutes, you may be able see bands in as little as five minutes, depending on how much DNA is present. Letting it run too long will result in losing your lower molecular weight bands. If your application requires loading more than the recommended amount of DNA, use the post-staining protocol. You cant have a good western blot without first having a good SDS gel. To avoid any interference the dye may have on DNA migration, we recommend using the post-staining protocol. This usually solves band migration problems. For samples of unknown concentration, try loading one half or one third of the usual amount of DNA. The technique uses three elements to accomplish this task: (1) separation by size, (2) transfer to a solid support, and (3) marking. By using a western blot, researchers are able to identify specific proteins from a complex mixture of proteins extracted from cells. The recommended loading amount for ladders and samples of known concentration is 50-200 ng/lane. Western blotting is an important technique used in cell and molecular biology. If you see band migration shifts or smearing and smiling, try reducing the amount of DNA loaded. Smearing and smiling in GelRed® or GelGreen® precast gels most often caused by overloading of DNA. Some samples, such as restriction digested DNA may migrate abnormally in GelRed® or GelGreen® precast gels. However, because GelRed® and GelGreen® are high affinity dyes designed to be larger dyes to improve their safety, they can affect the migration of DNA in precast gels. Many customers use GelRed® or GelGreen® precast gels for convenience. ![]()
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