A scalable method for multiplex LED-controlled synthesis of DNA in capillaries.
As research in synthetic biology and genomic sciences becomes more widespread, the need for diverse oligonucleotide populations has increased. To limit reagent cost, it would be advantageous to obtain high quality populations in minute amounts. Towards that end, synthesis of DNA strands in capillaries utilizing photolabile 3-nitrophenylpropyloxycarbonyl (NPPOC) chemistry and ultraviolet-light emitting diodes (UV-LEDs) was examined. Multiple oligonucleotides were made in single capillaries and were characterized by hybridization, sequencing and gene synthesis. DNA synthesized in capillaries was capable of being hybridized and signal intensities correlated with microarray data. Sequencing demonstrated that the oligonucleotides were of high quality (up to 44% perfect sequences). Oligonucleotides were combined and used successfully for gene synthesis. This system offers a novel, scalable method to synthesize high quality oligonucleotides for biological applications.
Blair
Sarah
S
Materials Science Program, University of Wisconsin, Madison, WI 53706, USA.
Richmond
Kathryn
K
Rodesch
Matthew
M
Bassetti
Michael
M
Cerrina
Franco
F
D002264:Carboxylic Acids / Q000737:chemistry; D004247:DNA / Q000138:chemical synthesis*; D005813:Genes, Synthetic; D005898:Glass / Q000737:chemistry; D009578:Nitrobenzenes / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis*; D017422:Sequence Analysis, DNA; D017640:Silicates / Q000737:chemistry; D014466:Ultraviolet Rays
C430114:2-nitrophenylpropyloxycarbonyl chloride; D002264:Carboxylic Acids; D009578:Nitrobenzenes; D009838:Oligodeoxyribonucleotides; D017640:Silicates; D004247:DNA