RADOM, an efficient in vivo method for assembling designed DNA fragments up to 10 kb long in saccharomyces cerevisiae

Qiuhui Lin, Bin Jia, Leslie A. Mitchell, Jingchuan Luo, Kun Yang, Karen I. Zeller, Wenqian Zhang, Zhuwei Xu, Giovanni Stracquadanio, Joel S. Bader, Jef D. Boeke, Ying Jin Yuan*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)213-220
Number of pages8
JournalACS Synthetic Biology
Volume4
Issue number3
DOIs
Publication statusPublished - 13 May 2014

Keywords

  • Build-A-Genome
  • in vivo assembly
  • Sc2.0
  • synthetic biology
  • synthetic yeast genome

Fingerprint

Dive into the research topics of 'RADOM, an efficient in vivo method for assembling designed DNA fragments up to 10 kb long in saccharomyces cerevisiae'. Together they form a unique fingerprint.

Cite this