[Oligonucleotide microarray preparation using enhanced poly-L-lysine glass slides].
OBJECTIVE
To modify conventional poly-L-lysine coating for oligonucleotide microarray preparation so as to enhance the sensitivity of the microarray.


METHODS
The proposed chemical approaches included silanizing the slides with 3-glycid-oxypropyltrimethoxysilane (GOPS) after cleaning, followed by slide coating with polymers (poly-L-lysine) that was covalently bound to the modified glass. Subsequent attachment of the oligonucleotide to the modified slide surface was achieved after 1,4-phenylene diisothiocyanate (PDITC) activation of the surface. Various experiments were carried out, such as the immobilization efficiency and hybridization assays to test the modified slides, which were then used tentatively in the preparation of microarrays for SARS coronavirus detection.


RESULTS
The improved surface had high immobilization efficiency, good uniformity and satisfactory hybridization efficiency, better than those slides with conventional poly-L-lysine coating. In addition, such modified slides rendered the microarrays more resistant to consecutive probing/stripping cycles.


CONCLUSIONS
The modified slide surface is satisfactory to immobilize unmodified oligonucleotide by covalent binding, which enhances not only the sensitivity of the prepared oligonucleotide microarray but also the binding of the oligonucleotide to the slide surface.
Wu
Qing-hua
QH
Institute of Molecular Biology, Southern Medical University, Guangzhou 510515, China. wuqh@fimmu.com
Ma
Wen-li
WL
Shi
Rong
R
Guo
Qiu-ye
QY
Zhang
Bao
B
Li
Ling
L
Zhang
Hai-yan
HY
Zheng
Wen-ling
WL
D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000592:standards*; D015345:Oligonucleotide Probes; D011107:Polylysine / Q000737:chemistry*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000302:isolation & purification*
D004279:DNA, Viral; D015345:Oligonucleotide Probes; D011107:Polylysine