Lin Wang, Yang Xi, Wenwei Zhang, Weimao Wang, Hanjie Shen, Xiaojue Wang, Xia Zhao, Andrei Alexeev, Brock A Peters, Alayna Albert, Xu Xu, Han Ren, Ou Wang, Killeen Kirkconnell, Helena Perazich, Sonya Clark, Evan Hurowitz, Ao Chen, Xun Xu, Radoje Drmanac, Yuan Jiang
Nucleic acid ligases are crucial enzymes that repair breaks in DNA or RNA during synthesis, repair and recombination. Various genomic tools have been developed using the diverse activities of DNA/RNA ligases. Herein, we demonstrate a non-conventional ability of T4 DNA ligase to insert 5' phosphorylated blunt-end double-stranded DNA to DNA breaks at 3'-recessive ends, gaps, or nicks to form a Y-shaped 3'-branch structure. Therefore, this base pairing-independent ligation is termed 3'-branch ligation (3'BL). In an extensive study of optimal ligation conditions, the presence of 10% PEG-8000 in the ligation buffer significantly increased ligation efficiency to more than 80%...
November 13, 2018: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes