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Article

PCR-based accurate synthesis of long DNA sequences.

Nature protocols - 1 Jan 2006

Xiong Ai-Sheng, Yao Quan-Hong, Peng Ri-He, Duan Hui, Li Xian, Fan Hui-Qin, Cheng Zong-Ming, Li Yi

Abstract excerpt

Here we describe a simple and rapid method for assembly and PCR-based accurate synthesis (PAS) of long DNA sequences. The PAS protocol involves the following five steps: (i) design of the DNA sequence to be synthesized and of 60-bp overlapping oligonucleotides to cover the entire DNA sequence; (ii) purification of the oligonucleotides by PAGE; (iii) first PCR, to synthesize DNA fragments of 400-500 bp in length...

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PMID: 17406309
DOI: 10.1038/nprot.2006.103

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