separation problem with GelRed

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Post by Biotium » Wed Nov 26, 2014 4:43 pm

I know this is a very old thread, but we just wanted to chime in with an official reply and answers to your questions.

GelRed is designed to be a large dye to increase its safety. Therefore, its large size may affect DNA migration in prestain gels, leading to some separation problems.

Here are some solutions to improve band separation:

1. Reduce the amount of loaded DNA. Overloading of DNA often results in band smearing. The recommended amount for ladders and samples of known concentration is 50-200 ng/lane. If the concentration is unknown, try loading one half or one third of the usual amount of DNA.
2. Reduce the amount of GelRed. Try using 0.5X instead of 1X final concentration of GelRed.
3. Pour a lower percentage of agarose gel. Higher molecular weight DNA separates better with a lower percentage gel.
4. Change the running buffer. TBE buffer has a higher buffering capacity than TAE buffer.
5. Try using the post-staining protocol if possible, especially if more than the recommended amount of DNA is required.

For other questions regarding GelRed, check out Biotium’s FAQ ... el-stains/
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