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New Methods for Generating and Studying Modified Porphyrins and Olignucleotides

Kendra Lee Dombi

出版	Tufts University, 2002
URL	http://books.google.com.hk/books?id=GKnMNwAACAAJ&hl=&source=gbs_api

註釋

In the third part of this thesis, a method for preparing small DNA-arrays on aldehyde-bearing glass slides is presented. The immobilization of the DNA involves reductive amination and employs oligonucleotides with 3' -terminal lysine residues, obtained in high yield from solid phase syntheses. Spot patterns are produced by protecting selected areas of the aldehyde-bearing slides with wax, coating the free surface with a methyl triethylene glycol derivative, and removing the wax with dichloromethane. The DNA arrays give better signal-to-noise ratios in hybridization experiments than slides without passified background. A model chip with the 5'-modified sequence, chl-T*GGTTGACTGCGAT-DP-Lys, where chl stands for a cholic acid residue, DP for a dimethylhydroxypropionic acid residue, and T* for a 5' -amino-5'deoxythymidine residue, was prepared and was shown to bind its target strand with higher affinity than its unmodified counterpart TGGTTGACTGCGAT-DP-Lys. The methodology for preparing background-passified oligonucleotide arrays is now being used to develop 'high fidelity DNA chips' whose mismatch discrimination is better than that of conventional chips.