Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 2448-2452 |
| Number of pages | 5 |
| Journal | ChemBioChem |
| Volume | 11 |
| Issue number | 17 |
| DOIs | |
| Publication status | Published - 2010 Nov 1 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Medicine
- Molecular Biology
- Organic Chemistry
Cite this
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A Fluorescence Selection Method for Accurate Large-Gene Synthesis. / Kim, Hwangbeom; Han, Hyojun; Shin, Dongyun; Bang, Duhee.
In: ChemBioChem, Vol. 11, No. 17, 01.11.2010, p. 2448-2452.Research output: Contribution to journal › Article
TY - JOUR
T1 - A Fluorescence Selection Method for Accurate Large-Gene Synthesis
AU - Kim, Hwangbeom
AU - Han, Hyojun
AU - Shin, Dongyun
AU - Bang, Duhee
PY - 2010/11/1
Y1 - 2010/11/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=78449255034&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=78449255034&partnerID=8YFLogxK
U2 - 10.1002/cbic.201000368
DO - 10.1002/cbic.201000368
M3 - Article
C2 - 20981747
AN - SCOPUS:78449255034
VL - 11
SP - 2448
EP - 2452
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 17
ER -