Facile Splint-Free Circularization of ssDNA with T4 DNA Ligase by Redesigning the Linear Substrate to Form an Intramolecular Dynamic Nick
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Circularization of Linear ssDNA
2.3. Evaluation of Circularization Yield and Selectivity
2.4. Mfold Calculation
3. Results
3.1. Unexpected Ligation of an Intramolecular Nick Flanking Only Two orThree Base Pairs to Circularize a 39-nt ssDNA
3.2. The Length Limit for Efficient PPC
3.3. Effect of Nick Location on Circularization
3.4. Optimize Reaction Conditions to Increase the Yield of Circularization by Decreasing or Avoiding Adenylation
3.5. Universality of the Method
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sun, W.; Hu, K.; Liu, M.; Luo, J.; An, R.; Liang, X. Facile Splint-Free Circularization of ssDNA with T4 DNA Ligase by Redesigning the Linear Substrate to Form an Intramolecular Dynamic Nick. Biomolecules 2024, 14, 1027. https://doi.org/10.3390/biom14081027
Sun W, Hu K, Liu M, Luo J, An R, Liang X. Facile Splint-Free Circularization of ssDNA with T4 DNA Ligase by Redesigning the Linear Substrate to Form an Intramolecular Dynamic Nick. Biomolecules. 2024; 14(8):1027. https://doi.org/10.3390/biom14081027
Chicago/Turabian StyleSun, Wenhua, Kunling Hu, Mengqin Liu, Jian Luo, Ran An, and Xingguo Liang. 2024. "Facile Splint-Free Circularization of ssDNA with T4 DNA Ligase by Redesigning the Linear Substrate to Form an Intramolecular Dynamic Nick" Biomolecules 14, no. 8: 1027. https://doi.org/10.3390/biom14081027