Gel for short DNA

Agarose/polyacrilamide composite gel is also called AgPAGE. In case of resolving DNA fragments use the same instruments as for standard agarose gel electrophoresis. In first place mix all components of PAGE except TEMED, then add warm agarose, mix well then add TEMED. The important moment – which temperature the agarose should have, because high temperature induces PA polymerization. If you are not intended to do gradient gel, i.e. you need just the gel, so I think you can use more hot agarose may be 40 – 50C. If you use low melting agarose you should decrease the temperature to about 32-35C. After addition of TEMED allow the gel to polymerize for about 1 hour at room temperature. May be you need optimize the concentration of the agarose and PA to obtain the better resolution. I think agarose should be at least 3.5% and PA at least 15% and may be more. In some protocols there are additional steps of treatment of the mix gel I think this is to make additional cross-links between agarose and PA


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