Figure 1. Stabilization of the hairpin structure by non-nucleotide linkers (-L-) following enzymatic circularization. Structures of the stilbene diether.

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Figure 1. Stabilization of the hairpin structure by non-nucleotide linkers (-L-) following enzymatic circularization. Structures of the stilbene diether and hexaethylene glycol (EG6) linkers are depicted. From: Stabilization of RNA hairpins using non-nucleotide linkers and circularization Nucleic Acids Res. 2017;45(10):e92. doi:10.1093/nar/gkx122 Nucleic Acids Res | © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

Figure 2. Circularization of RNA using T4 RNA ligase Figure 2. Circularization of RNA using T4 RNA ligase. (A) Schematic representation of RNA substrates: three oligomers containing CUG repeats and a sarcin–ricin (SR–G) sequence from Escherichia coli 23S rRNA. (B) Control (C) and ligation reactions (L) resolved on an 8% polyacrylamide gel in denaturing conditions. Open and circular forms of the RNA oligomers are indicated. From: Stabilization of RNA hairpins using non-nucleotide linkers and circularization Nucleic Acids Res. 2017;45(10):e92. doi:10.1093/nar/gkx122 Nucleic Acids Res | © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

Figure 3. Identification of the circular form of the RNA oligomer Figure 3. Identification of the circular form of the RNA oligomer. (A) Circularization reaction was carried out in the absence of ATP or T4 RNA ligase and separated on an 8% polyacrylamide gel. C—RNA oligomer only. (B) Autoradiogram shows separation of the products of alkaline hydrolysis to the linear and circular forms of the RNA oligomer (R). C—control reaction. (C) Identification of the circular RNA form using dephosphorylation. The reaction products were separated on an 8% polyacrylamide gel and visualized by autoradiography followed by staining with SYBR Green II. From: Stabilization of RNA hairpins using non-nucleotide linkers and circularization Nucleic Acids Res. 2017;45(10):e92. doi:10.1093/nar/gkx122 Nucleic Acids Res | © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

Figure 4. (A) Circularization efficiency of RNA using recombinant (rec Figure 4. (A) Circularization efficiency of RNA using recombinant (rec.) or commercially available enzymes (lanes A and B). The ligation efficiency is not affected by the annealing temperature (B) but depends on the ligation temperature. (C) Open and circular forms of the RNA oligomer are indicated. From: Stabilization of RNA hairpins using non-nucleotide linkers and circularization Nucleic Acids Res. 2017;45(10):e92. doi:10.1093/nar/gkx122 Nucleic Acids Res | © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

Figure 5. The normalized UV melting curves of the open (orange) and circular (blue) form of the 8G RNA oligomer. From: Stabilization of RNA hairpins using non-nucleotide linkers and circularization Nucleic Acids Res. 2017;45(10):e92. doi:10.1093/nar/gkx122 Nucleic Acids Res | © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

Figure 6. Stabilization of the hairpin structure by the click reaction Figure 6. Stabilization of the hairpin structure by the click reaction. Red and blue triangles represent the 5΄ and 3΄ end modifications required for circularization using the click reaction. The triazole linkage is depicted. From: Stabilization of RNA hairpins using non-nucleotide linkers and circularization Nucleic Acids Res. 2017;45(10):e92. doi:10.1093/nar/gkx122 Nucleic Acids Res | © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

Figure 7. Circularization of DNA and RNA oligomers using the click chemistry. (A) Separation of the click reaction products (R) along with the linear form of the oligomer (C). (B) Schematic representation of the open and closed forms of DNA oligomer. The HpaII restriction sites and length of the digestion products are marked. (C) 20% polyacrylamide gel electrophoresis (PAGE) of the restriction products of open and circular forms of the DNA oligomer. C—control lane. From: Stabilization of RNA hairpins using non-nucleotide linkers and circularization Nucleic Acids Res. 2017;45(10):e92. doi:10.1093/nar/gkx122 Nucleic Acids Res | © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com