Figure 2
From: RNase H-assisted RNA-primed rolling circle amplification for targeted RNA sequence detection
Comparison of the sealing efficiency of the RNA-splinted padlock probe under different ATP concentrations. (A) Denatured polyacrylamide gel analysis of products ligated by T4 DNA ligase (T4Dnl), T4 RNA ligase 2 (T4Rnl2), and SplintR ligase (SplintR) using an RNA-splinted padlock probe. The numbers above each lane indicate the ATP concentration (µM) used in the ligation reaction. M, 20-bp DNA size marker; Linear, linear padlock probe; Circular, circular padlock probe made by CircLigase. The white arrow indicates a linear padlock probe. The black arrow indicates a circularized padlock probe. The red arrows indicate oligo debris of DNase treatment. The blue arrow indicates a circularized padlock probe/RNA hybrid molecule. (B) Relative yield of the circularized padlock probe. The gel images were analyzed using ImageJ for comparisons with the known concentration of the pre-circularized probe made by CircLigase. The yield of each padlock probe was calculated by setting the control as 100%. Circ., circular probe made by CircLigase; Blank, blank lane. Five electrophoresis experiments were conducted using five independent reaction products, and the mean and error were calculated.
