A flexible light-directed DNA chip synthesis gated by deprotection using solution photogenerated acids.
Oligonucleotide microarrays or oDNA chips are effective decoding and analytical tools for genomic sequences and are useful for a broad range of applications. Therefore, it is desirable to have synthesis methods of DNA chips that are highly flexible in sequence design and provide high quality and general adoptability. We report herein, DNA microarray synthesis based on a flexible biochip method. Our method simply uses photogenerated acid (PGA) in solution to trigger deprotection of the 5'-OH group in conventional nucleotide phosphoramidite monomers (i.e. PGA-gated deprotection), with the rest of the reactions in the synthesis cycle the same as those used for routine synthesis of oligonucleotides. The complete DNA chip synthesis process is accomplished on a regular DNA synthesizer that is coupled with a UV-VIS projection display unit for performing digital photolithography. Using this method, oDNA chips containing probes of newly discovered genes can be quickly and easily synthesized at high yields in a conventional laboratory setting. Furthermore, the PGA-gated chemistry should be applicable to microarray syntheses of a variety of combinatorial molecules, such as peptides and organic molecules.
Gao
X
X
Department of Chemistry, University of Houston, Houston, TX 77004-5003, USA. xgao@uh.edu
LeProust
E
E
Zhang
H
H
Srivannavit
O
O
Gulari
E
E
Yu
P
P
Nishiguchi
C
C
Xiang
Q
Q
Zhou
X
X
D004247:DNA / Q000138:chemical synthesis* / Q000235:genetics; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000235:genetics; D010777:Photochemistry / Q000379:methods*; D016133:Polymerase Chain Reaction; D012996:Solutions
D009841:Oligonucleotides; D012996:Solutions; D004247:DNA