Computationally Optimised DNA Assembly of synthetic genes.
Gene synthesis is hampered by two obstacles: improper assembly of oligonucleotides; oligonucleotide defects incurred during chemical synthesis. To overcome the first problem, we describe the employment of a Computationally Optimised DNA Assembly (CODA) algorithm that uses the degeneracy of the genetic code to design overlapping oligonucleotides with thermodynamic properties for self-assembly into a single, linear, DNA product. To address the second problem, we describe a hierarchical assembly strategy that reduces the incorporation of defective oligonucleotides into full-length gene constructs. The CODA algorithm and these biological methods enable fast, simple and reliable assemblies of sequence-correct full-length genes.
Larsen
Liza S Z
LS
CODA Genomics Inc., Laguna Hills, CA 92653, USA. Larsen@cantab.net
Wassman
Christopher D
CD
Hatfield
G Wesley
GW
Lathrop
Richard H
RH
D000465:Algorithms; D001483:Base Sequence; D003198:Computer Simulation; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D005813:Genes, Synthetic / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D008956:Models, Chemical; D008957:Models, Genetic; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods*
D004247:DNA