Temperature conditions on casting a gel minimizing the band distortion of DGGE

Abstract

For denaturing gradient gel electrophoresis (DGGE) analysis, the band distortion has been known to be affected by the unevenness of an electric current and the buffer temperature during DNA loading. However, little is known about the effect of the temperature on the band distortion. The effect of the temperatures of the gel solution (denaturing gel solution temperature: DGT) and glass plates (plate temperature: PT) at the gel-pouring time was examined in the range of 5-28℃ and 10-28℃, respectively. The results showed that the band distortion increased as the difference between DGT and PT increased when the same DNA solutions were loaded in all wells. From these results, the combination of DGT and PT was considered to be a crucial factor for reducing the band distortion. These results and the study of the relationship between the gel polymerization rate and the combination of the two temperatures in an in vitro experiment suggested the heterogeneity of the polymerization rate within a glass plate.