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. Author manuscript; available in PMC: 2023 May 5.
Published in final edited form as: Mol Cell. 2022 Mar 30;82(9):1768–1777.e3. doi: 10.1016/j.molcel.2022.03.008

Figure 1. Circular RNA migration using standard agarose systems.

Figure 1.

(A) Schematic and expected sizes of precursor linear RNA, linear control RNA, circular RNA, and excised introns. Linear control RNA is splicing incompetent as it lacks the full upstream and downstream introns necessary for splicing but retains remnant introns expected after splicing. Probes targeting the indicated segments of the RNA are numbered 1–6 in the indicated colors.

(B) Northern blot correctly identifies circular RNA using standard agarose electrophoresis. Circular RNA and linear control RNA were treated with RNase R for 15 minutes and 60 minutes. Samples were incubated in NorthernMax-Gly Sample Loading Dye containing glyoxal, glycerol and DMSO, and run at 50V for 80 minutes at room temperature using traditional agarose gels cast in our lab. Size markers are indicated on the left; biotinylated probes used for each Northern blot are indicated on the left and correspond with panel A. RNA species are labeled on the right. Gel and Northern blot images were cropped above the 2000nt marker for clarity.