Development of an oligonucleotide based sandwich array platform for the detection of transgenic elements from plant sources using labal-free PCR products

Download

index.pdf

Date

2010

Author

Gül, Fatma

Metadata

Show full item record

Item Usage Stats

197
views

65
downloads

Advances in DNA micro and macroarray technologies made these high-throughput systems good candidates for the development of cheaper, faster and easier qualitative and quantitative detection methods. In this study, a simple and cost effective sandwich hybridization-based method has been developed for the rapid and sensitive detection of various unmodified recombinant elements in transgenic plants. Attention was first focused on the optimization of conditions such as time, concentration and temperature using commercial ssDNA, which in turn could be used for real sample detection. In this sandwich-type DNA chip platform, capture probes complementary to the first half of recombinant element (target adapter) were immobilized onto poly-L-lysine covered conventional microscope slides. PCR-amplified un-purified target adapter and biotin labeled detection probe, which is complementary to the second half of target adapter, were hybridized in solution-phase to complementary capture probes to create a sandwiched tripartite complex. Later, hybridization signal was visualized by the attachment of streptavidin conjugated Quantum Dot to the sandwiched complex under UV illumination. Sandwich based array system that has been developed in this study allows multiplex screening of GMO events on a single DNA chip platform. 35S promoter, NOS terminator, CRY1Ab and BAR target sequences were successfully detected on the same DNA chip platform. The platform was able to detect unlabeled PCR amplified DNA fragments of CaMV 35S promoter sequence and NOS terminator and BAR transgene sequences from transgenic potato plants and NK603 Certified GMO Reference material, respectively. The DNA-chip platform developed in this study will allow multiple detection of label-free PCR-amplified transgenic elements from real GMO samples on a single slide via a cost effective, fast, reliable and sensitive sandwich hybridization assay.

Subject Keywords

Technique.

URI

http://etd.lib.metu.edu.tr/upload/12612659/index.pdf
https://hdl.handle.net/11511/19937

Collections

Graduate School of Natural and Applied Sciences, Thesis

Suggestions

OpenMETU
Core

Development of a sandwich-type DNA array platform for the detection of label-free oligonucleotide targets Cansız, Sena; Öktem, Hüseyin Avni; Özen, Can; Department of Biotechnology (2010) DNA arrays have become a major bioanalytical method as they enable high-throughput screening and they can be manufactured on different surfaces depending on the nature of diagnostic purpose. However, current technologies to produce and detect arrays of DNA probes are expensive due to the requirement of specialized instrumentation. In this study we have established an array platform in sandwich hybridization format for the detection of label-free nucleic acid targets. Unlike direct hybridization, which is th...
Development of algorithms and software for forecasting, nowcasting and variability of TEC Tulunay, E; Senalp, ET; Cander, LR; Tulunay, YK; Bilge, AH; Mizrahi, E; Kouris, SS; Jakowski, N (2004-01-01) Total Electron Content (TEC) is an important characteristic of the ionosphere relevant to communications. Unpredictable variability of the ionospheric parameters due to various disturbances limits the efficiencies of communications, radar and navigation systems. Therefore forecasting and nowcasting of TEC are important in the planning and operation of Earth-space and satellite-to-satellite communication systems. Near-Earth space processes are complex being highly nonlinear and time varying with random varia...
Development of a regression model for the life assessment of open-hole specimens with double through cracks utilizing stress intensity factor calculations via XFEM Shabestari, S. Sohrab Heidari; Kayran, Altan (Elsevier BV; 2019-01-01) Development of a regression model for the life assessment of open-hole specimens with double through cracks utilizing stress intensity factor calculations via XFEM
Search for Pair Production of a Heavy Up-Type Quark Decaying to a W Boson and a b Quark in the lepton plus jets Channel with the ATLAS Detector Aad, G.; et. al. (2012-06-26) A search is presented for production of a heavy up-type quark (t') together with its antiparticle, assuming subsequent decay to a W boson and a b quark, t'(t) over bar' -> W(+)bW(-)(b) over bar. The search is based on 1.04 fb(-1) of proton-proton collisions at root s = 7 TeV collected by the ATLAS detector at the CERN Large Hadron Collider. Data are analyzed in the lepton + jets final state, characterized by a high transverse momentum isolated electron or muon, high missing transverse momentum, and at least...
Measurement of the Isolated Prompt Photon Production Cross Section in pp Collisions at root s=7 TeV Khachatryan, V.; et. al. (2011-02-01) The differential cross section for the inclusive production of isolated prompt photons has been measured as a function of the photon transverse energy E-T(Gamma) in pp collisions at root s 7 TeV using data recorded by the CMS detector at the LHC. The data sample corresponds to an integrated luminosity of 2.9 pb(-1). Photons are required to have a pseudorapidity vertical bar eta(gamma)vertical bar 21 GeV, covering the kinematic region 0.006 < x(T) < 0.086. The measured cross section is found to be in agreem...

Citation Formats

F. Gül, “Development of an oligonucleotide based sandwich array platform for the detection of transgenic elements from plant sources using labal-free PCR products,” M.S. - Master of Science, Middle East Technical University, 2010.