Pulsed Field Gel Electrophoresis (PFGE) System for the Separation of Large DNA Molecules
Rotating field electrophoresis (ROFE)
Pulsed field electrophoresis with the Rotaphor system allows the separation of DNA beyond the 50 kb limit of conventional agarose electrophoresis. Thanks to the patented electrode rotor, the electrical field can be applied in virtually any angle. To achieve a very homogeneous field, the two main rotor electrodes are flanked by 2 sets of secondary electrodes each.
Due to the unique design of the Rotaphor system, all common techniques for separation of large DNA molecules can be applied. This includes methods like CHEF, FIGE, PAGE and naturally ROFE (Rotating Field Electrophoresis).
Since PFGE gels typically run for many hours (up to several days), the electrophoresis buffer has to be cooled. The Rotaphor electrophoresis chamber comes with a built-in buffer circulation pump which is connected to an external cooling thermostat. During the run the buffer temperature in the electrophoresis chamber is constantly monitored and precisely controlled by the Rotaphor software.
The Rotaphor system includes a PC that controls the electrode rotor and the power supply over the Rotaphor interface card. The Rotaphor 8 software provides 17 pre-set programs for separation of different size ranges. Starting with the pre-set parameters, new applications are quickly optimised. For each pre-set program the software shows a real gel image. Once a custom program has been optimised, a referring gel picture can be uploaded into the software. Thus, the Rotaphor software over time will not only be a control tool, but also a library of successful experiments linked to the underlying separation parameters. By linking programs to combined lists also complex sequences can be programmed.