PCR electrophoresis tank EHS1 series

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The range of fragment sizes to be separated will determine the choice of agarose concentration for a gel. Typical agarose concentration is 0.5% to 3.0%. For large DNA fragments low-percentage gels are required, while for small DNA fragments, high-percentage gels are recommended. Weak gels (0.5% agarose) should be electrophoresed at low temperatures (e.g. -4°C). Agarose gels of 0.75% to 1.0%, for routine electrophoresis, are recommended for a wide range of separations (0.15 to15 kb). 2…4% agarose gels are usually selected for PCR fragment resolution. If the gel has to be subsequently photographed, thin gels (2 to 3 mm) with low-percentage agarose are better than thick or high-percentage gels. The latter produce increased opaqueness and autofluorescence. Electrophoresis buffer TAE buffer provides optimal resolution of fragments >4 kb in length, while for 0.1 to 3 kb fragments, TBE buffer should be selected. TBE has both a higher buffering capacity and lower conductivity than TAE and therefore should be used for high-voltage electrophoresis. Additionally, TBE buffer generates less heat than TAE at an equivalent voltage and does not allow a significant pH drift. Note: because of its lower buffering capacity, TAE should be circulated or mixed from time to time for full-length electrophoresis, especially at higher voltages. Temperature influence Electrophoresis at high voltages produces heat. Additionally, high-conductivity buffers such as TAE generate more heat than low-conductivity buffers.