Using a Temperature Gradient against the Time in Polyacrylamide Gel Electrophoresis May Eliminate the Need for Stacking Gels

Background and objectives: making stacking gels for polyacrylamide gels in the laboratoryby conventional methods is laborious and time consuming. considering the role of temperature inpolyacrylamide gels with respect to electrical resistance and viscosity, we assumed that decreasingthe temperature would cause an increase in electrical resistance and viscosity. ultimately, adownward temperature gradient imposed in the first phase of polyacrylamide gel running timewould supposedly improve the migration of macromolecules. this project analyzed the effect oftemperature gradient on the migration of macromolecules in the continuous gels (without stacking)and compared it with results obtained using stacking gels.material and methods: electrical resistance was calculated using ohm’s law. subsequently, toexamine the effect of temperature change on macromolecules separation, conformation sensitivegel electrophoresis (csge) was used as a model and specimens were run under three differentconditions, one of which was prepared with a stacking gel.results: the electrical resistance showed an inverse relationship with temperature in this study.separation of the dna molecules in the continuous gels (with no stacking) was comparable with theconventional method (with stacking).conclusion: using a temperature gradient against time may be an alternative method forstacking gels.