Ultra‐fast simultaneous analysis of genetically modified organisms in maize by microchip electrophoresis with LIF detector

Abstract

This study examined the potential of microchip electrophoresis (ME) with a LIF detector using a programmed field strength gradient (PFSG) in a conventional glass double-T microchip for the ultra-fast detection and simultaneous analysis of genetically modified (GM) maize. The separation efficiency and sensitivity at various sieving gels (poly(ethylene oxide) (PEO, M r 8 000 000) and 2-hydroxyethylcellulose (HEC) (M r 250 000)) and fluorescent dye concentrations were investigated. The PCR products of both the GM and non-GM maize were analyzed within 30 s under the PFSG (470.6 V/cm for 20 s, 117.6 V/cm for 12 s, and 470.6 V/cm for 30 s) with a 2.5% HEC sieving matrix in the running buffer, 16Tris-borate EDTA (TBE) (pH 8.30) and 0.5 ppm ethidium bromide. The five transgenic maize varieties (Event176, MON810, Bt11, GA21, and T25) examined in this study were also clearly differentiated by ME-PFSG within 30 s in a single run without any loss of resolution. The ME-PFSG technique is a powerful tool for the ultra-fast detection and simultaneous analysis of GMOs in a variety of foods including maize.

Bioanalytical separations using electric field gradient techniques

“…The effects of varying buffer concentrations and electric field strengths on the separation of 202 and 273 bp DNA fragments were analyzed, and the PFSG performed almost seven times faster than microchip-based capillary gel A few other ME-PFSG studies investigated combining the system with an LIF detector. PCR products of genetically modified and non-genetically modified maize varieties were examined by Kumar and Kang [110]. Results demonstrated that a 30 s run could discriminate between five genetically modified maize varieties; this short analysis time is a vast improvement (up to 60 times) over earlier published data [132].…”

“…Five papers utilized programmed field strength gradients (PFSG) to improve electrophoretic separations [110][111][112][113][114], while the remainder featured a novel or improved DEP device capable of capturing or manipulating DNA [18,68,[115][116][117][118][119][120][121][122][123][124][125][126][127][128]. The PFSG studies focus on detection techniques of various PCR products, whereas the DEP papers centered on the manipulation, trapping, sorting, or electrostretching of various-sized DNA.…”

Establishment of Dipstick Development Technology for Detection of Cry1Ac in Transgenic Plants

“…These include: the optimum pH of the reaction, the appropriate concentration of protein loaded onto the colloids, determining the isoelectric point (pI) of the protein, and the stability of the colloids. Generally, most proteins can be adsorbed onto the metal surface in an optimal manner using buffer systems close to their pI value, away from this pH the adsorption decreases [16]. Gold nano particles were being used in current study as color producing probes because these are stated to be non-toxic, inert and have long retention of their optical properties, which creates them a better choice as a signal generator.…”

On-line capillary electrophoresis for enhanced detection sensitivity of feline panleukopenia virus

“…To reduce analysis time, a programmed field strength gradient (PFSG) technique has been performed [13][14][15][16][17]. PFSG can control the electrophoretic velocity of DNA molecules by changing electric fields.…”