Rapid hybridization kinetics of DNA attached to submkron latex particles

Wolf, Stanley F.; Haines, Lora L.; Fisch, Judith E.; Kremsky, Jonathan N.; Dougherty, Joseph P.; Jacobs, Kenneth

doi:10.1093/nar/15.7.2911

Cited by 99 publications

(47 citation statements)

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“…New approaches to differential hybridization to identify rare mRNA clones from cDNA libraries using single-stranded cDNA (Boll etal., 1986); 5. Use of UV-irradiation to covalently bind DNA to solid supports (Stollar and Rashtchian, 1987) and immobilization of DNA to Sephacryl 500 dextran supports, which improve hybridization efficiencies to 80% compared to 20% with filter hybridization (Goldkorn and Prockop, 1986;Gingeras, Kwoh and Davis, 1987;Thompson and Gillespie, 1987;Wolf et al, 1987); 6. Hybridization strategies that reduce background from samples or enable the use of solution hybridization, which can reduce hybridization times to a few minutes (Gingeras, Kwoh and Davies, 1987;Karjalainen, Rouhainen and Soderlund, 1987;Nicholls, Langdale and Malcolm, 1987;Wilkins and Snell, 1987;Palva, Nyberg and Palva, 1988;Urdea etal., 1988); 7.…”

Section: Nucleic Acid Hybridization Methodsmentioning

confidence: 99%

New Approaches to the Detection of Microbial Plant Pathogens

Chu

Waterhouse

Martín

et al. 1989

Biotechnology and Genetic Engineering Reviews

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Section: Nucleic Acid Hybridization Methodsmentioning

confidence: 99%

New Approaches to the Detection of Microbial Plant Pathogens

Chu

Waterhouse

Martín

et al. 1989

Biotechnology and Genetic Engineering Reviews

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“…NucleoLink strips were selected for the present study because the binding efficiency of DNA to NucleoLink has been shown to be five to 10 times higher than to CovaLink (Oroskar et al, 1996). NucleoLink strips are made of an activated heat-stable polymer and support covalent binding of DNA by carbodiimide-mediated condensation, and these strips can be assembled in micro-well format and used in micro-well plate readers (Chu et al, 1983 ;Ghosh & Musso, 1987 ;Oroskar et al, 1996 ;Rasmussen et al, 1991 ;Wolf et al, 1987).…”

Section: Introductionmentioning

confidence: 99%

DNA-DNA hybridization determined in micro-wells using covalent attachment of DNA.

Christensen¹,

Angen²,

Mutters³

et al. 2000

International Journal of Systematic and Evolutionary Microbiology

173

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The present study was aimed at reducing the time and labour used to perform DNA-DNA hybridizations for classification of bacteria at the species level. A micro-well-format DNA hybridization method was developed and validated. DNA extractions were performed by a small-scale method and DNA was sheared mechanically into fragments of between 400 and 700 bases. The hybridization conditions were calibrated according to DNA similarities obtained by the spectrophotometric method using strains within the family Pasteurellaceae. Optimal conditions were obtained with 300 ng DNA added per well and bound by covalent attachment to NucleoLink. Hybridization was performed with 500 ng DNA, 5 % (w/w) of which was labelled with photoactivatable biotin (competitive hybridization) for 25 h at 65 SC in 2iSSC followed by stringent washing with 2iSSC at the same temperature. The criteria for acceptance of results were a maximum of 15 % standard deviation, calculated as a percentage of the mean for four replicate micro-wells, and that DNA similarities were not significantly different in at least two independent experiments. The relationship between DNA similarities obtained by the microwell method (y) and by the spectrophotometric method (x) was y l 0534xM306, when these criteria had been applied to 23 pairs of strains of Actinobacillus species, avian [Pasteurella] haemolytica-like bacteria and Mannheimia species. The correlation (Pearson) between DNA similarities obtained by interchange of strains used for covalent binding and hybridization was 0794. Significantly lower DNA similarities were observed by the spectrophotometric compared with the micro-well method for three pairs of hybridizations. After removal of these data, the relationship between DNA similarities obtained by the micro-well and spectrophotometric methods improved to y l 0855xM110. It was found that the accuracy and precision of the micro-well method was at the same level as that of the spectrophotometric method, but the labour and analysis time were reduced significantly. The use of hybridization in the micro-well format will allow DNA-DNA hybridizations to be carried out between all strains selected for a particular taxonomic study, in order to construct complete data matrices and improve species definition.

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“…Wolf et al compared rates for hybridization of a radiolabeled oligomer to DNA-coated particles with solution-phase hybridization rates and demonstrated that rates on particles approached those in solution (ϳ0.2x-1x) for the conditions tested (33). Later Jungell-Nortamo and co-workers determined that capture of radiolabeled nucleic acid hybrids onto plasmid-coated microparticles, although somewhat slower than solution-phase hybridization, was significantly faster than capture onto membrane-immobilized plasmid (34).…”

mentioning

confidence: 99%