Kim, Hwangbeom Han, Hyojun Shin, Dongyun Bang, Duhee 2024-01-20T18:04:12Z 2024-01-20T18:04:12Z 2021-09-05 2010-11-22 1439-4227 https://pubs.kist.re.kr/handle/201004/130907 The fundamental problem for low-cost gene synthesis is errors that occur during the synthetic process. To address this problem, we developed a practical method that exploits the fact that the predominant errors are deletions. In this method, a simple fluorescence-based readout was used to distinguish error-free synthetic DNA molecules. To do this, we constructed vectors that contained multiple cloning sites and GFP. In the vectors, the GFP gene is designed to be out-of-frame, but insertion of an in-framed synthetic DNA construct into the appropriate cloning site will lead to fluorescent cell colonies. We successfully used this method to synthesize five genes and improved the bp per error from 629 to 6552 by selecting green fluorescent colonies. English WILEY-V C H VERLAG GMBH DNA EXPRESSION IDENTIFICATION SEQUENCES CLONING GENOME A Fluorescence Selection Method for Accurate Large-Gene Synthesis Article 10.1002/cbic.201000368 1 CHEMBIOCHEM, v.11, no.17, pp.2448 - 2452 CHEMBIOCHEM 11 17 2448 2452 scie scopus 000284718100016 2-s2.0-78449255034 Biochemistry & Molecular Biology Chemistry, Medicinal Biochemistry & Molecular Biology Pharmacology & Pharmacy Article DNA EXPRESSION IDENTIFICATION SEQUENCES CLONING GENOME polymerase chain reaction synthetic biology DNA synthesis gene technology