ROMJIST Volume 20, No. 4, 2017, pp. 426-439
M. Banu, M. Simion, P. Varasteanu, L. Savu, I. Farcasanu Microarray and Surface Plasmon Resonance experiments for HPV genotyping on Au-supports
ABSTRACT: This paper is focused on obtaining DNA-modified gold biochips by ensuring the hybridization conditions of DNA and establishing the optimal co-immobilization ratios of DNA/ mercaptohexanol (MCH). Microarray technique and statistical analysis were employed to assess the immobilization protocol of random DNA sequences. By considering two types of probe carbon-spacer (C spacer), HPV 18 oligonucleotides were tethered on gold support with or without MCH and the probe attachment kinetics were investigated by Surface Plasmon Resonance (SPR). Further, the hybridization conditions of 4 different concentrations of real HPV-18 fragments were studied. The probe immobilization efficiency was achieved after 2 h of incubation using buffer without MCH, or with DNA: MCH concentrations of 1:1 and 1:2, whereas the fluorescently labelled DNA and the probes with C6 spacer revealed superior hybridization capabilities. Also, SPR characterization disclosed promising hybridization results by using real DNA fragments corresponding to HPV 18.KEYWORDS: MCH, co-immobilization, hybridization, C spacer, HPV 18, microarray, SPRRead full text (pdf)