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. 2024 Jun;30(6):739–747. doi: 10.1261/rna.079895.123

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© 2024 Zhang et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article, published in RNA, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.

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FIGURE 3. — Optimization of TL-qPCR efficiency regarding m¹A proximity and ligation temperature. (A) Oligonucleotides used and ligation oligo design. (B) Amplification curves of linker 5lkrT/C/G/A in A and m¹A oligo in ligation. (C) Ligation oligo design at varying ligation sites away from the m¹A site. (D) Cq plots as a function of ligation oligos with ligation sites at m¹A (T + 0) or progressively away from the m¹A site. (E) ΔCq values between the m¹A site and control site (using average Cq values of sites ≥4 nt away from the m¹A site) under different template splint ligation temperatures.