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RADOM, an Efficient <i>In Vivo</i> Method for Assembling Designed DNA Fragments up to 10 kb Long in <i>Saccharomyces cerevisiae</i>

Lin, Qiuhui and Jia, Bin and Mitchell, Leslie A and Luo, Jingchuan and Yang, Kun and Zeller, Karen I and Zhang, Wenqian and Xu, Zhuwei and Stracquadanio, Giovanni and Bader, Joel S and Boeke, Jef D and Yuan, Ying-Jin (2015) 'RADOM, an Efficient <i>In Vivo</i> Method for Assembling Designed DNA Fragments up to 10 kb Long in <i>Saccharomyces cerevisiae</i>.' ACS Synthetic Biology, 4 (3). pp. 213-220. ISSN 2161-5063

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We describe rapid assembly of DNA overlapping multifragments (RADOM), an improved assembly method via homologous recombination in Saccharomyces cerevisiae, which combines assembly in yeasto with blue/white screening in Escherichia coli. We show that RADOM can successfully assemble ∼3 and ∼10 kb DNA fragments that are highly similar to the yeast genome rapidly and accurately. This method was tested in the Build-A-Genome course by undergraduate students, where 125 ∼3 kb "minichunks" from the synthetic yeast genome project Sc2.0 were assembled. Here, 122 out of 125 minichunks achieved insertions with correct sizes, and 102 minichunks were sequenced verified. As this method reduces the time-consuming and labor-intensive efforts of yeast assembly by improving the screening efficiency for correct assemblies, it may find routine applications in the construction of DNA fragments, especially in hierarchical assembly projects.

Item Type: Article
Uncontrolled Keywords: in vivo assembly; synthetic yeast genome; Sc2.0; Build-A-Genome; synthetic biology
Subjects: Q Science > QA Mathematics > QA75 Electronic computers. Computer science
Q Science > QD Chemistry
Divisions: Faculty of Science and Health
Faculty of Science and Health > Computer Science and Electronic Engineering, School of
SWORD Depositor: Elements
Depositing User: Elements
Date Deposited: 16 Jan 2017 12:44
Last Modified: 18 Aug 2022 11:32

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