Design of UltraHigh Density DNALaden Polylysinemodified 设计超高密度聚赖氨酸修饰DNA拉登.pptVIP

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Design of UltraHigh Density DNALaden Polylysinemodified 设计超高密度聚赖氨酸修饰DNA拉登.ppt

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Design of UltraHigh Density DNALaden Polylysinemodified 设计超高密度聚赖氨酸修饰DNA拉登

Design of Ultra-High Density DNA-Laden Polylysine-modified Silicon Surfaces Presented by Michael D. Soriano Graduate Mentor: Yuli Wang Faculty Mentor: Dr. Ying C. Chang UC LEADS Program August 15, 2002 Abstract Single-stranded DNA was attached to polylysine-modified silicon (PLS) surfaces. Initially, the PLS substrate was protected. Using a mixture of HBr/CH3COOH, the surface was deprotected, ready for the coupling reaction with the natural DNA, 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDAC), and N-hydroxysuccinimide (NHS). Measurements of the characterization of each surface were taken: the contact angle, refractive index, and thickness before and after deprotection; thickness and refractive index after the coupling reactions. The coupling reactions were optimized by modifying the surface density of the PLS. Binding specificity of complementary DNA was tested for using fluorescence hybridization. Preliminary results showed increases in substrate thickness from 20-80% after the coupling reactions. Introduction The current technology of DNA microarrays attaches the nucleotides via bi-hetero-functional linkers1 or heat/light2, e.g. ultraviolet (UV) light. Protocols for microarray design call for coating silicon or glass slides with polylysine (PLL), then DNA attachment is mediated. The resulting microarrays may effectively only be used once and have a superficial coating of DNA on the surfaces. Lenigk et al.3 have created microarrays for repeated use by using (3-mercaptopropyl)trimethoxysilane. They have successfully shown that their design may be used repeatedly up to five times. Our design implements attachment of DNA strands to polylysine substrates. We have effectively increased the thickness of the substrate from 20 – 80% after coupling reactions. Background fluorescence contaminated the sample Materials/MethodsDeprotection of Substrates Protected substrates according to Wang4 Substrates were suspended in a layer of benze