Immobilization of DNA in Polyacrylamide Gel for the Manufacture of DNA and DNA–Oligonucleotide Microchips

Proudnikov, Dmitri; Timofeev, Edward N.; Mirzabekov, Andrei D.

doi:10.1006/abio.1998.2620

Cited by 170 publications

(114 citation statements)

References 22 publications

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“…On other substrates, increasing surface area (by applying acrylamide gels to glass slides, for example) to allow the immobilization of larger amounts of DNA resulted in greater signal intensities and an increased dynamic range. [35] We took advantage of the high surface area of the paper by using a paper with a relatively high profile root mean square roughness parameter, R R.M.S. of 6.4 ± 1.9 µm, area root mean square roughness parameter S R.M.S ., of 10.7 ± 0.6 µm, and the porosity, or the volume fraction of void, of ~68%.…”

Section: Supportmentioning

confidence: 99%

Analytical Devices Based on Direct Synthesis of DNA on Paper

et al. 2015

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This paper addresses a growing need in clinical diagnostics for parallel, multiplex analysis of biomarkers from small biological samples. It describes a new procedure for assembling microarrays of ssDNA and proteins on paper. This method starts with the synthesis of DNA oligonucleotides covalently linked to paper, and proceeds to generate DNA arrays capable of simultaneously capturing DNA, DNA-conjugated protein antigens, and DNA-conjugated antibodies. The synthesis of ssDNA oligonucleotides on paper is convenient and effective, with32% of the oligonucleotides cleaved and eluted from the paper substrate being full-length by HPLC for a 32-mer. These ssDNA arrays can be used to detect fluorophore-linked DNA oligonucleotides in solution, and as the basis for DNA-directed assembly of microarrays of DNA-conjugated capture antibodies on paper, detect protein antigens by sandwich ELISAs. Paper-anchored ssDNA arrays with different sequences can be used to assemble paper-based devices capable of detecting DNA and antibodies in the same device, and enable simple microfluidic paperbased devices.3

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Section: Supportmentioning

confidence: 99%

Analytical Devices Based on Direct Synthesis of DNA on Paper

et al. 2015

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“…It is similar to a PCR-based microarray except that no amplification is required. Another type of array has also been fabricated by immobilizing oligonucleotides in a polyacrylamide gel (Proudnikov et al, 1998).…”

Section: Oligonucleotide-based Dna Microarraysmentioning

confidence: 99%

“…However, the cost of making long oligonucleotides is high. There are a limited number of whole microbial genome arrays that are available in the Affymetrix system (Proudnikov et al, 1998).…”

Section: Oligonucleotide-based Dna Microarraysmentioning

confidence: 99%

DNA microarrays and their applications in medical microbiology

Nsofor¹

2014

Biotechnol. Mol. Biol. Rev.

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Rapid diagnosis and treatment of disease is often based on the identification and characterization of causative agents derived from phenotypic characteristics. This can be laborious and time consuming, often requiring many skilled personnel and a large amount of lab space. However, the introduction of nucleic acid amplification techniques into molecular biology has transformed the laboratory detection of pathogens. The progression of the molecular diagnostic revolution currently relies on the ability to efficiently and accurately offer multiplex detection and characterization for a variety of infectious disease pathogens. DNA microarray analysis has the capability to offer robust multiplex detection. Multiple microarray platforms exist, including printed double-stranded DNA and oligonucleotide arrays, in situ-synthesized arrays, high-density bead arrays, electronic microarrays, and suspension bead arrays. The aim of this paper was to review DNA microarray technology, highlighting two major types: the oligonucleotide-based array and the PCR product-based array. Although, the use of microarrays to generate gene expression data has become routine, applications pertinent to microbiology continue to rapidly expand. This review highlights uses of microarray technology that impact diagnostic microbiology, including the detection and identification of pathogens, determination of antimicrobial resistance, epidemiological strain typing, and determination of virulence factors.

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“…7). A photopolymerization process described previously was used to create the array of gel pads (Proudnikov et al, 1998), after which the glass slide was treated in 0.1M NaBH 4 for 30 min to reduce gel pads auto-fluorescence. The Biochip Arrayer (PerkinElmer, Boston, MA) was used to spot beads onto singular gel pads on the device.…”

Section: Wwwintechopencommentioning

confidence: 99%