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Comparison between Different Strategies of Covalent Attachment of DNA to Glass Surfaces to Build DNA Microarrays

✍ Scribed by Nathalie Zammatteo; Laurent Jeanmart; Sandrine Hamels; Stéphane Courtois; Pierre Louette; Laszlo Hevesi; José Remacle

Publisher
Elsevier Science
Year
2000
Tongue
English
Weight
186 KB
Volume
280
Category
Article
ISSN
0003-2697

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✦ Synopsis

DNA microarray is a powerful tool allowing simultaneous detection of many different target molecules present in a sample. The efficiency of the array depends mainly on the sequence of the capture probes and the way they are attached to the support. The coupling procedure must be quick, covalent, and reproducible in order to be compatible with automatic spotting devices dispensing tiny drops of liquids on the surface. We compared several coupling strategies currently used to covalently graft DNA onto a glass surface. The results indicate that fixation of aminated DNA to an aldehyde-modified surface is a choice method to build DNA microarrays. Both the coupling procedure and the hybridization efficiency have been optimized. The detection limit of human cytomegalovirus target DNA amplicons on such DNA microarrays has been estimated to be 0.01 nM by fluorescent detection.

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