DNA microarrays on a dendron-modified surface improve significantly the detection of single nucleotide variations in the p53 gene.
Selectivity and sensitivity in the detection of single nucleotide polymorphisms (SNPs) are among most important attributes to determine the performance of DNA microarrays. We previously reported the generation of a novel mesospaced surface prepared by applying dendron molecules on the solid surface. DNA microarrays that were fabricated on the dendron-modified surface exhibited outstanding performance for the detection of single nucleotide variation in the synthetic oligonucleotide DNA. DNA microarrays on the dendron-modified surface were subjected to the detection of single nucleotide variations in the exons 5-8 of the p53 gene in genomic DNAs from cancer cell lines. DNA microarrays on the dendron-modified surface clearly discriminated single nucleotide variations in hotspot codons with high selectivity and sensitivity. The ratio between the fluorescence intensity of perfectly matched duplexes and that of single nucleotide mismatched duplexes was >5-100 without sacrificing signal intensity. Our results showed that the outstanding performance of DNA microarrays fabricated on the dendron-modified surface is strongly related to novel properties of the dendron molecule, which has the conical structure allowing mesospacing between the capture probes. Our microarrays on the dendron-modified surface can reduce the steric hindrance not only between the solid surface and target DNA, but also among immobilized capture probes enabling the hybridization process on the surface to be very effective. Our DNA microarrays on the dendron-modified surface could be applied to various analyses that require accurate detection of SNPs.
Oh
Soon Jin
SJ
Division of Molecular and Life Sciences, Pohang University of Science and Technology Pohang 790-784, Korea.
Ju
Jimin
J
Kim
Byung Chul
BC
Ko
Eunsil
E
Hong
Bong Jin
BJ
Park
Jae-Gahb
JG
Park
Joon Won
JW
Choi
Kwan Yong
KY
D000873:Anthracenes / Q000737:chemistry*; D045744:Cell Line, Tumor; D015342:DNA Probes / Q000737:chemistry; D005453:Fluorescence; D016158:Genes, p53; D006579:Heterozygote; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D020641:Polymorphism, Single Nucleotide
D000873:Anthracenes; D015342:DNA Probes; C501244:dendron