DNA microarrays with PAMAM dendritic linker systems.
The DNA microarray-based analysis of single nucleotide polymorphisms (SNPs) is important for the correlation of genetic variations and individual phenotypes, and for locating disease-causing genes. To facilitate the development of surfaces suitable for immobilization of oligonucleotides, we report here a novel method for the surface immobilization of DNA using pre-fabricated polyamidoamine (PAMAM) starburst dendrimers as mediator moieties. Dendrimers containing 64 primary amino groups in their outer sphere are covalently attached to silylated glass supports and, subsequently, the dendritic macromolecules are modified with glutaric anhydride and activated with N-hydroxysuccinimide. As a result of the dendritic PAMAM linker system the surfaces reveal both a very high immobilization efficiency for amino-modified DNA-oligomers, and also a remarkable high stability during repeated regeneration and re-using cycles. The performance of dendrimer-based DNA microarrays in the discrimination of SNPs is demonstrated.
Benters
Rüdiger
R
Chimera Biotec GmbH, Schwachhauser Heerstrasse 30A, 28209 Bremen, Germany.
Niemeyer
Christof M
CM
Drutschmann
Denja
D
Blohm
Dietmar
D
Wöhrle
Dieter
D
D000812:Anhydrides / Q000737:chemistry; D020137:Base Pair Mismatch / Q000235:genetics; D003247:Conservation of Natural Resources; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis / Q000379:methods; D015342:DNA Probes / Q000235:genetics / Q000378:metabolism; D050091:Dendrimers; D005453:Fluorescence; D005898:Glass / Q000737:chemistry; D005977:Glutarates / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011073:Polyamines / Q000737:chemistry* / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012680:Sensitivity and Specificity; D013388:Succinimides / Q000737:chemistry
D000812:Anhydrides; D015342:DNA Probes; D050091:Dendrimers; D005977:Glutarates; C104700:PAMAM Starburst; D011073:Polyamines; D013388:Succinimides; C079643:glutaric anhydride; D004247:DNA; C001426:N-hydroxysuccinimide