title: Non-polymerase-cycling-assembly-based chemical gene synthesis: strategies, methods, and progress.
abstract: Chemical gene synthesis is a powerful tool for basic biological research and biotechnology applications. During the last 30 years, major advances have been made in the chemical synthesis of DNA sequences ranging from fragments of <1 kb to multi-gene sequences of >30 kb. There is a need for simple, reproducible, less error-prone and cost-effective methods that guarantee successful synthesis of the desired genes and are amenable to automation. Many polymerase chain reaction (PCR)-based and non-polymerase-cycling-assembly (PCA)-based strategies have been developed for chemical gene synthesis. The PCR-based method has been the subject of several recent reviews. Here, we provide an overview of the progress in non-PCA-based chemical gene synthesis using different strategies and methods, including enzymatic gene synthesis, annealing and ligation reaction, simultaneous synthesis of two genes via a hybrid gene, shotgun ligation and co-ligation, insertion gene synthesis, gene synthesis via one strand of DNA, template-directed ligation, ligase chain reaction, microarray-mediated gene synthesis, Blue Heron solid support technology and Sloning building block technology. The fundamental principle underlying each strategy, an example where applicable, and the advantages and disadvantages are discussed. The emphasis is on discussion of the most recent technologies and their potential applications, particularly for microarray-based genomics research.
authors: Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China.
authors: Ai-Sheng
authors: AS
authors: Xiong
authors: Ri-He
authors: RH
authors: Peng
authors: Jing
authors: J
authors: Zhuang
authors: Jin-Ge
authors: JG
authors: Liu
authors: Feng
authors: F
authors: Gao
authors: Jian-Min
authors: JM
authors: Chen
authors: Zong-Ming
authors: ZM
authors: Cheng
authors: Quan-Hong
authors: QH
authors: Yao
mesh_terms: D001709:Biotechnology / Q000379:methods* / Q000639:trends*; D004247:DNA / Q000138:chemical synthesis*; D005544:Forecasting; D005796:Genes; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000639:trends*; D016133:Polymerase Chain Reaction
chemical_list: D004247:DNA