title: DATEL: A Scarless and Sequence-Independent DNA Assembly Method Using Thermostable Exonucleases and Ligase.
abstract: DNA assembly is a pivotal technique in synthetic biology. Here, we report a scarless and sequence-independent DNA assembly method using thermal exonucleases (Taq and Pfu DNA polymerases) and Taq DNA ligase (DATEL). Under the optimized conditions, DATEL allows rapid assembly of 2-10 DNA fragments (1-2 h) with high accuracy (between 74 and 100%). Owing to the simple operation system with denaturation-annealing-cleavage-ligation temperature cycles in one tube, DATEL is expected to be a desirable choice for both manual and automated high-throughput assembly of DNA fragments, which will greatly facilitate the rapid progress of synthetic biology and metabolic engineering.
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, ‡The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, and §Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University , Wuxi 214122, P. R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: Jin
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, ‡The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, and §Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University , Wuxi 214122, P. R. China.
authors.forename: Wenwen
authors.initials: W
authors.lastname: Ding
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, ‡The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, and §Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University , Wuxi 214122, P. R. China.
authors.forename: Guocheng
authors.initials: G
authors.lastname: Du
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, ‡The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, and §Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University , Wuxi 214122, P. R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, ‡The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, and §Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University , Wuxi 214122, P. R. China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Kang
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D011088:DNA Ligases / Q000378:metabolism*; D005092:Exonucleases / Q000378:metabolism*; D058615:Synthetic Biology / Q000379:methods; D013696:Temperature
chemical_list: D004247:DNA; D005092:Exonucleases; D011088:DNA Ligases
keywords: DNA assembly; metabolic engineering; synthetic biology; thermostable exonuclease and ligase