Abstract
An improved and simplified protocol for DNA immobilization was developed to enhance DNA-DNA hybridization on microwell plates. Target DNA was immobilized by simple dry-adsorption. Efficiencies of DNA immobilization and retention were enhanced 1.4-6.5 times and 4.2-19.6 times, respectively, compared with a conventional method. The overall hybridization efficiency was increased 3.1-5.2 times. This simple new protocol can reduce the consumption of scarce DNA samples.
Full Text
The Full Text of this article is available as a PDF (69.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ezaki T., Takeuchi N., Liu S. L., Kai A., Yamamoto H., Yabuuchi E. Small-scale DNA preparation for rapid genetic identification of Campylobacter species without radioisotope. Microbiol Immunol. 1988;32(2):141–150. doi: 10.1111/j.1348-0421.1988.tb01373.x. [DOI] [PubMed] [Google Scholar]
- Forster A. C., McInnes J. L., Skingle D. C., Symons R. H. Non-radioactive hybridization probes prepared by the chemical labelling of DNA and RNA with a novel reagent, photobiotin. Nucleic Acids Res. 1985 Feb 11;13(3):745–761. doi: 10.1093/nar/13.3.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inouye S., Hondo R. Microplate hybridization of amplified viral DNA segment. J Clin Microbiol. 1990 Jun;28(6):1469–1472. doi: 10.1128/jcm.28.6.1469-1472.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lüneberg E., Jensen J. S., Frosch M. Detection of Mycoplasma pneumoniae by polymerase chain reaction and nonradioactive hybridization in microtiter plates. J Clin Microbiol. 1993 May;31(5):1088–1094. doi: 10.1128/jcm.31.5.1088-1094.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nagata Y., Yokota H., Kosuda O., Yokoo K., Takemura K., Kikuchi T. Quantification of picogram levels of specific DNA immobilized in microtiter wells. FEBS Lett. 1985 Apr 22;183(2):379–382. doi: 10.1016/0014-5793(85)80814-0. [DOI] [PubMed] [Google Scholar]
- Rasmussen S. R., Larsen M. R., Rasmussen S. E. Covalent immobilization of DNA onto polystyrene microwells: the molecules are only bound at the 5' end. Anal Biochem. 1991 Oct;198(1):138–142. doi: 10.1016/0003-2697(91)90518-x. [DOI] [PubMed] [Google Scholar]
- Vekris A., Bauduer F., Maillet S., Bébéar C., Bonnet J. Improved microplate immunoenzymatic assay of PCR products for rapid detection of Mycoplasma pneumoniae. Mol Cell Probes. 1995 Feb;9(1):25–31. doi: 10.1016/s0890-8508(95)90947-8. [DOI] [PubMed] [Google Scholar]
