Circular RNA migration in agarose gel electrophoresis - Abe

Correctly identifying circular RNA from its linear counterpart is essential to the RNA community. In this paper, we sought to determine how agarose gel electrophoresis conditions such as time, sample buffer, and voltage using the E-Gel EX system affect the migration of circular RNA compared to linear forms. We found that EDTA and use of SYBR-GOLD dye had the largest effect of slowing circular RNA migration, making it appear larger than its linearized form. We raise awareness of this variation in circular RNA migration and offer orthogonal methods in addition to gel electrophoresis such as northern blot, enzymatic assays, and HPLC to verify the RNA species.