DNA microarraying on compact disc surfaces. Application to the analysis of single nucleotide polymorphisms in Plum pox virus

Chemical Communications

Volumn , Issue 22, 2006, Pages 2368-2370

  Morais, Sergi ^a   Marco Molés, Raquel ^a   Puchades, Rosa ^a   Maquieira, Ángel ^a  

^a UNIVERSITAT POLITÈCNICA DE VALÈNCIA   (Spain)

Author keywords

[No Author keywords available]

Indexed keywords

NUCLEIC ACID; OLIGONUCLEOTIDE; PLANT DNA; POLYMER; PROTEIN;

ARTICLE; COMPACT DISK; DNA DETERMINATION; DNA MICROARRAY; GENETIC PROCEDURES; GENETIC VARIABILITY; MOLECULAR PROBE; MOSAIC VIRUS; NUCLEIC ACID ANALYSIS; NUCLEOTIDE SEQUENCE; POLYMERIZATION; PROTEIN ANALYSIS; SENSITIVITY ANALYSIS; SINGLE NUCLEOTIDE POLYMORPHISM; SURFACE PROPERTY; TECHNOLOGY; VIRAL GENETICS;

EID: 33744927449     PISSN: 13597345     EISSN: None     Source Type: Journal    
DOI: 10.1039/b600049e     Document Type: Article

References (23)

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12. This work was supported by the projects BIO2000-0243-P4-03 and BQU2003-02677 (CICYT, Spain)
13. DNA oligonucleotides were purchased from Tib Molbiol (Berlin, Germany)
14. Avidin (from egg white, extra-pure reagent) was from Sigma-Aldrich (Madrid, Spain). Buffers (PBS, 10 mM phosphate buffer, 150 mM NaCl, pH 7.4; PB, 150 mM phosphate buffer; CB, 50 mM carbonate-bicarbonate buffer, pH 9.6) were filtered through a 0.22 μm pore size disc before use. The polycarbonate reading faces of CD-Rs (PC) and PMMA slides (25 × 75 mm) were coated overnight at 4°C with 100 μL avidin (100 mg/L in CB). Then, surfaces were extensively washed with water and dried by slight centrifugation
15. PMMA slides, 1 mm thick, were kindly provided by Plexi S. A. (Valencia, Spain). CD-Rs were from U-Tech Media Corp., (Tau-Yuan Shien, Taiwan)
16. The fluorescence was measured adjusting the laser power and PMT settings at 100% and 600 V. The laser scanning system was from Axon Inst. (Union City, CA)
17. Contact angles were measured using an OCA 20 Video-Based Contact Angle Meter from DataPhysics Instruments, GmbH (Filderstadt, Germany). The thickness of the avidin film was measured using a Therma-Wave Opti-Probe 5220 (Fremont, CA)
18. The supporting information includes experimental aspects of the topology of coated surfaces and the influence of coating and printing buffer on fluorescence
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20. For these experiments, CB buffer as coating buffer and 3XSSC (450 mM NaCl, 45 mM sodium citrate, pH 7.0) as printing buffer were used as controls
21. Surfaces were incubated in hybridization chambers for 3 h at 42°C. After the hybridization step, the supports were first washed with 0.3XSSC and then with 0.1XSSC and water for 1 min. Hybridization buffer was 5XSSC containing 250 μg/mL sheared salmon sperm DNA, 0.02% BSA (w/v), and 25% formamide (v/v)
22. M. Cambra A. Olmos M. T. Gorris Bull. OEPP 2004 34 247
23. The authors wish to thank Dr. M. Cambra who kindly provided PPV primers and Dr. A. Olmos and Dra. N. Capote for helping in the design of probes for SNPs analysis