High-quality gene assembly directly from unpurified mixtures of microarray-synthesized oligonucleotides.

ABSTRACT: To meet the growing demand for synthetic genes more robust, scalable and inexpensive gene assembly technologies must be developed. Here, we present a protocol for high-quality gene assembly directly from low-cost marginal-quality microarray-synthesized oligonucleotides. Significantly, we eliminated the time- and money-consuming oligonucleotide purification steps through the use of hybridization-based selection embedded in the assembly process. The protocol was tested on mixtures of up to 2000 oligonucleotides eluted directly from microarrays obtained from three different chip manufacturers. These mixtures containing <5% perfect oligos, and were used directly for assembly of 27 test genes of different sizes. Gene quality was assessed by sequencing, and their activity was tested in coupled i

SUBMITTER: Borovkov AY 

PROVIDER: S-EPMC2965257 | biostudies-literature | 2010 Oct

REPOSITORIES: biostudies-literature