Effect of Charge Nature on the Adsorption of Single-Stranded DNA Fragments onto Latex Particles
✍ Scribed by A. Elaı̈ssari; J.P. Chauvet; M.A. Halle; O. Decavallas; C. Pichot; Ph. Cros
- Publisher
- Elsevier Science
- Year
- 1998
- Tongue
- English
- Weight
- 203 KB
- Volume
- 202
- Category
- Article
- ISSN
- 0021-9797
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✦ Synopsis
lose or nylon membranes. However, the slow kinetic rate The adsorption of single-stranded DNA fragments ( oligonuof the reaction between the nucleic acids in solution and cleotides ) onto latex particles is investigated using a series of those immobilized on the solid support is a drawback polymer latexes exhibiting various surface charge and group of this technique ( 1 ) . Polymeric microspheres such as natures ( sulfate, amidine, and amine ) . The latexes are prepolystyrene latex are of particular interest because they pared using free emulsion polymerization. In this work, we have been shown to exhibit good hybridization kinetics study the adsorption of single labeled oligonucleotide poly-( 1 ) . They are prepared easily at little cost with a wellthymidylic acid and of a 35-base-bearing fluorescein group defined size distribution and, furthermore, to permit a ( Fluo-dT 35 ) on four latexes to elucidate certain general princilarge variety of functions ( 2, 3 ) .
ples of oligonucleotide adsorption as a function of surface
Several methods for the covalent immobilization of sincharge nature. Several results are presented in this paper. It gle-stranded DNA fragments ( oligonucleotides, ODN ) on was found that the adsorbed amount of oligonucleotide is strongly dependent on the pH of the medium and the charge solid supports have been described ( 4 -7 ) and high levels density of positively charged latex particles. This indicates that of unspecified adsorption of the oligonucleotides have electrostatic forces play a major role in the adsorption process. been reported ( 7 ) . The adsorption of large fragments of The maximal adsorbed amount was found to be lower than the deoxyribonucleic acid and denatured calf thymus DNA one calculated from the jamming limit for the random sequenonto negatively charged ( sulfate ) polystyrene latex were tial adsorption of nonoverlapping rigid rods adsorbed in two first reported by Yamaoka et al. ( 8 ) . Several studies have dimensions. Desorption of the oligonucleotide was also studied been reported recently by on the by fluorescence measurements, in which both the supernatant adsorption of oligonucleotides on both cationic and anand the latex particles were analyzed using the front face analyionic latexes. It has been surmised that electrostatic forces sis method. In addition, attempts were made to correlate the play a major role in the adsorption process ( 12, 13 ) .
maximum fluorescence emission intensity using the front face
The adsorbed amount has been discussed by taking into analysis with the amount of oligonucleotides adsorbed onto account the effect of pH, ionic strength, and oligonucleolatex particles. ᭧ 1998 Academic Press tide nature and chain length ( 14,15 ) . Indeed, the effect