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  • Following hybridization on sensor/array platforms by using SPR, elipsometer and MALDI-MS.

Following hybridization on sensor/array platforms by using SPR, elipsometer and MALDI-MS.

Nucleosides, nucleotides & nucleic acids (2020-05-14)
Ömür Çelikbıçak, Kadriye Özlem Hamaloğlu, Bekir Salih, Erhan Pişkin
ABSTRACT

The aim of this study is to develop a methodology in which Surface Plasmon Resonance (SPR), Ellipsometer (EM) and Matrix-Assisted Laser Desorption/Ionization-Mass Spectrometry (MALDI-MS) will be used together for detection of single-strand oligodeoxynucleotides (ssODNs) targets. A selected target-ssODNs, and its complementary, the probe-ssODNs carrying a -SH end group, a spacer arm (HS-(CH2)6-(T)15, and a non-complementary ssODNs were used. Silicone based stamps with 16 regions were prepared and used for micro-contact printing (µCP) of the probe-ssODNs on the gold coated surfaces homogeneously. A modulator-spacer molecule (6-mercapto-1-hexanol) was co-immobilized to control surface probe density, to orientate the probe-ssODNs, and to eliminate the nonspecific interactions. SPR was used successfully to follow the hybridization of the target-ssODNs with the immobilized probe-ssODNs on the platform surfaces. Complete hybridizations were achieved in 100 min. It was obtained that there was a linear relationship between relative change in delta and target concentration below 1 µm. Using imaging version of ellipsometer (IEM) allowed imaging of the surfaces and supported extra datum for the SPR results. After a very simple dehybridization protocol, MALDI-MS analysis allowed detection of the target-ssODNs hybridized on the sensor/array platforms.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
1,3-Bis(N-carbazolyl)benzene, 97%
Sigma-Aldrich
Poly(dimethylsiloxane), viscosity 1.0 cSt (25 °C)