Microfluidic PicoArray synthesis of oligodeoxynucleotides and simultaneous assembling of multiple DNA sequences. Export

@article{citeulike:2874866, abstract = {Large DNA constructs of arbitrary sequences can currently be assembled with relative ease by joining short synthetic oligodeoxynucleotides (oligonucleotides). The ability to mass produce these synthetic genes readily will have a significant impact on research in biology and medicine. Presently, high-throughput gene synthesis is unlikely, due to the limits of oligonucleotide synthesis. We describe a microfluidic PicoArray method for the simultaneous synthesis and purification of oligonucleotides that are designed for multiplex gene synthesis. Given the demand for highly pure oligonucleotides in gene synthesis processes, we used a model to improve key reaction steps in DNA synthesis. The oligonucleotides obtained were successfully used in ligation under thermal cycling conditions to generate DNA constructs of several hundreds of base pairs. Protein expression using the gene thus synthesized was demonstrated. We used a DNA assembly strategy, i.e. ligation followed by fusion PCR, and achieved effective assembling of up to 10 kb DNA constructs. These results illustrate the potential of microfluidics-based ultra-fast oligonucleotide parallel synthesis as an enabling tool for modern synthetic biology applications, such as the construction of genome-scale molecular clones and cell-free large scale protein expression.}, address = {Department of Chemistry, University of Houston, Houston, TX 77004-5003, USA.}, author = {Zhou, X. and Cai, S. and Hong, A. and You, Q. and Yu, P. and Sheng, N. and Srivannavit, O. and Muranjan, S. and Rouillard, J. M. and Xia, Y. and Zhang, X. and Xiang, Q. and Ganesh, R. and Zhu, Q. and Matejko, A. and Gulari, E. and Gao, X.}, citeulike-article-id = {2874866}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/15477391}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=15477391}, issn = {1362-4962}, journal = {Nucleic acids research}, keywords = {biology, synthesis, synthetic}, number = {18}, pages = {5409--5417}, posted-at = {2008-06-09 09:37:28}, priority = {2}, title = {Microfluidic PicoArray synthesis of oligodeoxynucleotides and simultaneous assembling of multiple DNA sequences.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/15477391}, volume = {32}, year = {2004} }

TY - JOUR ID - citeulike:2874866 L3 - citeulike-article-id:2874866 N2 - Large DNA constructs of arbitrary sequences can currently be assembled with relative ease by joining short synthetic oligodeoxynucleotides (oligonucleotides). The ability to mass produce these synthetic genes readily will have a significant impact on research in biology and medicine. Presently, high-throughput gene synthesis is unlikely, due to the limits of oligonucleotide synthesis. We describe a microfluidic PicoArray method for the simultaneous synthesis and purification of oligonucleotides that are designed for multiplex gene synthesis. Given the demand for highly pure oligonucleotides in gene synthesis processes, we used a model to improve key reaction steps in DNA synthesis. The oligonucleotides obtained were successfully used in ligation under thermal cycling conditions to generate DNA constructs of several hundreds of base pairs. Protein expression using the gene thus synthesized was demonstrated. We used a DNA assembly strategy, i.e. ligation followed by fusion PCR, and achieved effective assembling of up to 10 kb DNA constructs. These results illustrate the potential of microfluidics-based ultra-fast oligonucleotide parallel synthesis as an enabling tool for modern synthetic biology applications, such as the construction of genome-scale molecular clones and cell-free large scale protein expression. IS - 18 JF - Nucleic acids research SN - 1362-4962 AD - Department of Chemistry, University of Houston, Houston, TX 77004-5003, USA. EP - 5417 TI - Microfluidic PicoArray synthesis of oligodeoxynucleotides and simultaneous assembling of multiple DNA sequences. VL - 32 SP - 5409 KW - biology KW - synthesis KW - synthetic AU - Zhou, X AU - Cai, S AU - Hong, A AU - You, Q AU - Yu, P AU - Sheng, N AU - Srivannavit, O AU - Muranjan, S AU - Rouillard, JM AU - Xia, Y AU - Zhang, X AU - Xiang, Q AU - Ganesh, R AU - Zhu, Q AU - Matejko, A AU - Gulari, E AU - Gao, X PY - 2004/// UR - http://view.ncbi.nlm.nih.gov/pubmed/15477391 ER -