Combinational synthesis of oligonucleotides and assembly fabrication of oligonucleotide array.
In this paper, a simple, reliable and flexible method, which integrated in situ synthesis with the spotting technique, was reported to fabricate oligonucleotide array. Different oligonucleotide sequences are synthesized on their relative code glass slides through combinational chemistry, thus the slides are broken into smaller pieces, in which the same code pieces have the same probe sequences. An oligonucleotide array is fabricated by arbitrarily assembling these different code pieces onto another solid substrate. In principle experimentation, four different sequences of P16 gene were synthesized and a 5 x 5 array including these four sequences and the control black was fabricated. The analysis results indicated that the hybridization fluorescence intensity of the same sequences locating different sets on the array gave the approximate values, and the fluorescence intensity ratio of matched sequence to one middle location base mismatched, two base mismatched, three middle base mismatched is (1.000+/-0.080):(0.4991+/-0.0671):(0.2360+/-0.0044):(0.0493+/-0.0033). Their relative accuracies were from 6.64 to 10.2%. This result might be used to rapidly screen single-nucleotide polymorphisms (SNPs).
Xiao
P F
PF
Chien-Shiung Wu Laboratory, Department of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China. xiaopf@seu.edu.cn
Wang
Z F
ZF
Guo
H S
HS
He
N Y
NY
Lu
Z H
ZH
D002232:Carbocyanines; D020650:Combinatorial Chemistry Techniques; D015335:Molecular Probes; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000138:chemical synthesis*
D002232:Carbocyanines; D015335:Molecular Probes; D009841:Oligonucleotides; C089187:cyanine dye 3