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An efficient covalent coating on glass slides for preparation of optical oligonucleotide microarrays
Atefeh Pourjahed1, Mohammad Rabiee2, Mohammadreza Tahriri2
1Biomaterials Group (Center of Excellence in Biomaterials), Faculty of Biomedical Engineering, Amirkabir University of Technology, Tehran, Iran ; Nanobiotechnology center, Department of Chemical Engineering, Jondishapour University of Technology, Dezful, Iran.
Objective(S):
Microarrays are potential analyzing tools for genomics and proteomics researches, which is in needed of suitable substrate for coating and also hybridization of biomolecules.
Materials And Methods:
In this research, a thin film of oxidized agarose was prepared on the glass slides which previously coated with poly-L-lysine (PLL). Some of the aldehyde groups of the activated agarose linked covalently to PLL amine groups; also bound to the amino groups of biomolecules. These linkages were fixed by UV irradiation. The prepared substrates were compared to only agarose-coated and PLL-coated slides.
Results:
RESULTS on atomic force microscope (AFM) demonstrated that agarose provided three-dimensional surface which had higher loading and bindig capacity for biomolecules than PLL-coated surface which had two-dimensional surface. In addition, the signal-to-noise ratio in hybridization reactions performed on the agarose-PLL coated substrates increased two fold and four fold compared to agarose and PLL coated substrates, respectively.
Conclusion:
The agarose-PLL microarrays had the highest signal (2546) and lowest background signal (205) in hybridization, suggesting that the prepared slides are suitable in analyzing wide concentration range of analytes.
