Oligonucleotides biotinylated

Lebaron, P. Catala, P. Fajon, C. Joux, F. Baudart, J. Bernard, L. A new sensitive, whole-cell hybridization technique for detection of bacteria involving a biotinylated oligonucleotide probe targeting rRNA and tyramide signal amplification. Appl. Environ. Microbiol. 1997, 63, 3274-3278. 

Amide bond linkage resulting in 5 -biotinylated oligonucleotide... 

Lloyd, R.V., Jin, L., and Fields, K. (1990) Detection of chromogranins A and B in endocrine tissues with radioactive and biotinylated oligonucleotide probes. Am. I. Surg. Pathol. 14, 35-43. 

P2 In this study, we report the chemical synthesis and functionalization of magnetic and gold-coated magnetic nanoparticles and the immobilization of single-stranded biotinylated oligonucleotides onto these particles. (From Kouassi and Irudayaraj, 2006)... 

Biotinylated proteins such as immunoglobulins and serum albumins or biotinylated oligonucleotides can bind through affinity interactions streptavidin-functiona-lized Au NPs [156, 157]. 

Amide Bond Linkage Resulting in 5 -Biotinylated Oligonucleotide... 

Immobilisation of oligonucleotide probes onto the electrode surface 40 pL of 3 -biotinylated oligonucleotide probes (0.5ng/pL) is left on the electrode surface for 15 min. Finally, the electrodes are rinsed with 2 x SSC buffer pH 7.2 containing 1% of BSA. 

Form the sensing phase placing 40 pL of S -biotinylated oligonucleotide probes (0.5ng/pL) for 15 min. 

Biotin is a common reagent for labelling oligonucleotides, and a new amidite (175) has been prepared for biotinylated oligonucleotides, but the biotin group may be removed by fluoride treatment.Biotinylated ddNTPs have been used for single base extension for multiplex genotyping by mass spectros-... 

As known from bioconjugate chemistry additional functional groups can be used for attachment of biomolecules. Thus, coupling via SH-groups [149,155,160], a benzaldehyde-semicarbazide-reaction [161] or the reaction between a streptavidin-coated surface and a biotinylated oligonucleotide [150,162,163] are only some examples. 

The LiPA control should show reaction on lines 1, 2, 3, 6, 11, 13, and 16. The LiPA control consists of biotinylated oligonucleotides complementary to the hybridization probes applied on these lines. Positive hybridization to these lines demonstrates satisfactory performance of the assay including hybridization, washing, and detection. The amplification control sample gives a clear positive signal on gel and on the strip on lines 1, 2, 3, 6,11,13, and 16. 

An alternative reagent (216) has been prepared for the synthesis of biotinylated oligonucleotides. In this case the protection used for the biotin moiety is base labile rather than the conventional dimethoxytrityl. The photo-crosslinking behaviour of oligonucleotide constructs, incorporating photoactive residues 217a-d at a defined position, has been examined in the presence of their DNA and RNA complementary targets. The X-ray crystal-structure of the photoproduct formed between 4-thiothymidine and adenosine upon near UV irradiation has been reported. 

Capture Biotinylated Oligonucleotides and Perform ELISA for Digoxigenin... 

The most common alternative for immobilization of oligonucleotides involves the use of biotinylated derivatives. These are conveniently made with standard reagents for oligonucleotide synthesis and biotin can be added at both the 3 and 5 ends. Immobilization of the biotinylated oligonucleotide to avidin-modified sensor surfaces is performed in neutral buffer conditions and is normally efficient. 

Ladd et al. reported SPR sensor-based detection of hCG [52], exploiting a DNA-directed antibody immobilization method. The immobilization consisted of non-covalent attachment of streptavidin to a biotinylated SAM followed by binding of biotinylated oligonucleotides to available streptavidin binding sites. Antibodies chemically modified with oligonucleotides with a complementary sequence were finally attached to this surface via DNA hybridization. The detection limit for direct detection of hCG in buffer by a dual-channel SPR sensor with wavelength modulation was determined to be... 

Figure 7.1. Schematic representation of different DNA immobilization strategies (a] covalent attachment of ODN thiols or disulfides by self-assembly onto gold surfaces resulting in Au-S bond formation (b) immobilization by adsorption relies on electrostatic interactions between negatively charged sugar-phosphate backbone of DNA and positively charged electrode surface, and/or the interaction with the nucleobases (c] affinity binding of biotinylated oligonucleotides onto streptavidin modified electrode surfaces.

Moreover, the significance of the attachment of biotinylated oligonucleotide probes through the streptavidin/biotin interaction has been tested in a previous work [15]. When a double-labeled (biotin and fluorescein) poly-T was attached to the electrode surface through the streptavidin/biotin interaction, the peak currents were much higher than those obtained when it was accumulated on the electrode surface by physical adsorption. This fact means that streptavidin/biotin interaction allows to attach and orient the oligonucleotide strands on electrode surface, whereas the direct adsorption of the oligonucleotide on the electrode surface results... 

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