10.1021/jo970872s.s001
Michael C. Pirrung
Michael C.
Pirrung
Lara Fallon
Lara
Fallon
Glenn McGall
Glenn
McGall
Proofing of Photolithographic DNA Synthesis with
3‘,5‘-Dimethoxybenzoinyloxycarbonyl-Protected Deoxynucleoside
Phosphoramidites
American Chemical Society
1998
probe
GTAGCATCTT
hybridization
nonpolar medium
homopolymers
target deoxyeicosanucleotide
Proofing
irradiation
sequence
method
solvent
efficiency
amidite
monomer
Photolithographic DNA Synthesis
nucleoside building blocks
phosphoramidite
cycle yields
fluorescein
pyrimidine
array
purine
Phosphoramidite
synthesis
microchip format
hybridized
covalently
photolabile
al
Photolysis rates
nm
Deprotection
McGall
GAC
derivatized glass surfaces
data
DMBOC
AAXTAXCTAC
Deoxynucleoside
16 decanucleotides
Surface fluorescence imaging
1998-01-01 00:00:00
Journal contribution
https://acs.figshare.com/articles/journal_contribution/Proofing_of_Photolithographic_DNA_Synthesis_with_3_5_-Dimethoxybenzoinyloxycarbonyl-Protected_Deoxynucleoside_Phosphoramidites/3707013
We have evaluated in a microchip format the photochemical
solid-phase phosphoramidite DNA
synthesis method we previously developed. A set of nucleoside
building blocks with “easy-off” base
protecting groups was prepared bearing photolabile
5‘-O-dimethoxybenzoincarbonate (DMBOC)
groups. Photolysis rates and cycle yields for these
DMBOC-protected nucleotides covalently attached
to planar, derivatized glass surfaces were determined by fluorescence
imaging-based methods earlier
developed by McGall et al. and described in detail elsewhere. Data were
obtained for both 280/310
and 365/400 nm irradiation in a range of solvents. Deprotection of
the DMBOC occurs fastest in
a nonpolar medium or without solvent. The coupling efficiency of
these amidites in the synthesis
of homopolymers was determined to be in the range 80−97%, with
purines generally showing lower
efficiency than pyrimidines. These DMBOC-protected monomers were
used to prepare a 4 × 4
array of 16 decanucleotides of the sequence 5‘-AAXTAXCTAC−chip, where
X = A, C, G, or T. The
array was hybridized with a target deoxyeicosanucleotide of the
sequence fluorescein-5‘-CTGAACG<b>GTAGCATCTT</b>GAC. Surface fluorescence imaging demonstrated
sequence-specific hybridization
to this probe.