One-step immobilization of aminated and thiolated DNA onto poly(methylmethacrylate) (PMMA) substrates

Fixe, F.; Dufva, Martin; Telleman, Pieter; Christensen, Claus Bo Vöge

doi:10.1039/b316616c

Cited by 102 publications

(114 citation statements)

References 33 publications

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“…However, the high autofluorescence of standard polymeric materials is not compatible with a highly sensitive detection method that employs fluorescent targets. Plastic microarray hybridizations compatible with fluorescent dyes have been developed previously (5,6,13). However, these investigators reported only hybridizations with labeled oligonucleotides, and the specific hybridization signal-to-background ratios were relatively low.…”

Section: Discussionmentioning

confidence: 99%

“…The only other reported study of room temperature hybridization on plastic supports was performed using a 16-h protocol, and the hybridization signal-to-background ratios obtained were low (approximately 10) (4). Others have performed hybridization experiments on plastic supports at 30 to 65°C, requiring special heating devices to carry out the hybridizations (5,6,15,17,23,25). In addition, stability tests demonstrated that when stored in a vacuum desiccator, the chemically activated plastic slides were stable for at least 10 weeks.…”

Section: Discussionmentioning

confidence: 99%

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Plastic Polymers for Efficient DNA Microarray Hybridization: Application to Microbiological Diagnostics

et al. 2008

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Fabrication of microarray devices using traditional glass slides is not easily adaptable to integration into microfluidic systems. There is thus a need for the development of polymeric materials showing a high hybridization signal-to-background ratio, enabling sensitive detection of microbial pathogens. We have developed such plastic supports suitable for highly sensitive DNA microarray hybridizations. The proof of concept of this microarray technology was done through the detection of four human respiratory viruses that were amplified and labeled with a fluorescent dye via a sensitive reverse transcriptase PCR (RT-PCR) assay. The performance of the microarray hybridization with plastic supports made of PMMA [poly(methylmethacrylate)]-VSUVT or Zeonor 1060R was compared to that with high-quality glass slide microarrays by using both passive and microfluidic hybridization systems. Specific hybridization signal-to-background ratios comparable to that obtained with high-quality commercial glass slides were achieved with both polymeric substrates. Microarray hybridizations demonstrated an analytical sensitivity equivalent to approximately 100 viral genome copies per RT-PCR, which is at least 100-fold higher than the sensitivities of previously reported DNA hybridizations on plastic supports. Testing of these plastic polymers using a microfluidic microarray hybridization platform also showed results that were comparable to those with glass supports. In conclusion, PMMA-VSUVT and Zeonor 1060R are both suitable for highly sensitive microarray hybridizations.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Plastic Polymers for Efficient DNA Microarray Hybridization: Application to Microbiological Diagnostics

et al. 2008

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“…34 In this case, the methyl ester groups at the surface of PMMA undergo a nucleophilic attack by the amine groups and this gives directly an amide linkage upon elimination of methanol. Although this method is simple, the authors not only indicate a high variability in their results, but Target #2 59-TAGGACACTCATAAT-39 59-Cy5 also report that the ''coupling reactions fail occasionally'' and this may be due to the high pH employed in the reaction as DNA may undergo hydrolysis.…”

Section: Surface Chemistry and Immobilization Of Dna On Poly(cyclic Omentioning

confidence: 99%

Surface modification of thermoplastics—towards the plastic biochip for high throughput screening devices

et al. 2007

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/npsi/ctrl?action=rtdoc&an=3539033&lang=en http://nparc.cisti-icist.nrc-cnrc.gc.ca/npsi/ctrl?action=rtdoc&an=3539033&lang=frAccess and use of this website and the material on it are subject to the Terms and Conditions set forth at http://nparc.cisti-icist.nrc-cnrc.gc.ca/npsi/jsp/nparc_cp.jsp?lang=en NRC Publications Archive Archives des publications du CNRCThis publication could be one of several versions: author's original, accepted manuscript or the publisher's version. / La version de cette publication peut être l'une des suivantes : la version prépublication de l'auteur, la version acceptée du manuscrit ou la version de l'éditeur. For the publisher's version, please access the DOI link below./ Pour consulter la version de l'éditeur, utilisez le lien DOI ci-dessous.http://dx.doi.org/10.1039/b700322fLab on Chip, 7, 2007-01-01 Microarrays have become one of the most convenient tools for high throughput screening, supporting major advances in genomics and proteomics. Other important applications can be found in medical diagnostics, detection of biothreats, drug discovery, etc. Integration of microarrays with microfluidic devices can be highly advantageous in terms of portability, shorter analysis time and lower consumption of expensive biological analytes. Since fabrication of microfluidic devices using traditional materials such as glass is rather expensive, there is great interest in employing polymeric materials as a low cost alternative that is suitable for mass production. A number of commercially available plastic materials were reviewed for this purpose and poly(methylmethacrylate) Zeonor 1060R and Zeonex E48R were identified as promising candidates, for which methods for surface modification and covalent immobilization of DNA oligonucleotides were developed. In addition, we present proof-of-concept plastic-based microarrays with and without integration with microfluidics.

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“…For the latter method, generally aminated or thiolated oligonucleotides are used [3][4][5]. The key problems to overcome for the preparation of DNA array based sensors or chips are the attachment of functional groups to the solid surface, the synthesis of oligonucleotides containing reactive groups, the spacial distribution of the DNA oligomers, and the detection of hybridization with complementary strands.…”

mentioning

confidence: 99%

Characterization and sequence verification of thiolated deoxyoligonucleotides used for microarray construction

Rozenski

2006

J. Am. Soc. Mass Spectrom.

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During synthesis of thiolated deoxyoligonucleotides, side products can be formed. When used in the fabrication of microchips, the oligonucleotides have to be of high purity. We demonstrated the presence of impurities, which were not failure sequences from the synthesis. These products were identified and characterized using high-performance liquid chromatography (HPLC) and electrospray MS(/MS). The presence of the free thiol group was assessed by acrylamide derivatization. After reaction with acrylamide the correct compounds showed a 71 u mass shift and the major fragment ions could be assigned as 5= a-base and 3= w ions, similar as for unmodified DNA. The side products were unaffected by acrylamide, suggesting that the thiol group was modified. The oligonucleotide containing a species with a mass of 32 u higher was identified as 5= sulfinic acid containing molecule and was found as 45% of the total amount of a DNA 25 mer. Other components appeared to be dithio-linked oxidation products, present about 1 to 5% in a 10 mer and 15 mer deoxyoligonucleotide. The analyses were useful for optimizing the synthesis

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One-step immobilization of aminated and thiolated DNA onto poly(methylmethacrylate) (PMMA) substrates

Cited by 102 publications

References 33 publications

Plastic Polymers for Efficient DNA Microarray Hybridization: Application to Microbiological Diagnostics

Plastic Polymers for Efficient DNA Microarray Hybridization: Application to Microbiological Diagnostics

Surface modification of thermoplastics—towards the plastic biochip for high throughput screening devices

Characterization and sequence verification of thiolated deoxyoligonucleotides used for microarray construction

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