Mark L. Collins

Nucleic acid hybridization assays employing dA-tailed capture probes. Single capture methods

Several novel hybridization techniques are described. Cells or specimens are treated to release nucleic acids and a liquid phase hybridization is carried out with a dA-tailed capture probe and a reporter probe in chaotropic salts or in salts containing SDS/proteinase K. In another format the tailed capture probe is preimmobilized on polystyrene and used to capture target nucleic acids from the solution. No phenol extraction or centrifugation is required to prepare the nucleic acids. Capture of the target on the poly (dT)-solid supports is used to remove excess labelled probe and sample impurities prior to non-radioisotopic or radioisotopic detection. This paper shows the advantage of a single round of capture on polystyrene, including the ability to assay large numbers of samples manually, the ability to analyse each sample for many analytes simultaneously, the use of rapid non-radioisotopic detection, and the ability to readily adapt the assay for automation.