From hybridization image to numerical values: a practical, high throughput quantification system for high density filter hybridizations

• Published in: Genetic analysis : biomolecular engineering Vol. 12; no. 3-4; p. 151
• Main Authors: Granjeaud, S, Nguyen, C, Rocha, D, Luton, R, Jordan, B R
• Format: Journal Article
• Language: English
• Published: Netherlands 01-01-1996
• Subjects: Animals; DNA, Complementary; Feasibility Studies; Filtration; Image Processing, Computer-Assisted; Mice; Nucleic Acid Hybridization; Peptide Elongation Factor 1; Peptide Elongation Factors - genetics; Reproducibility of Results; Software
• DOI: 10.1016/1050-3862(95)00128-X

Hybridization to sets of bacterial colonies or PCR products arrayed on high density filters is used in a number of experimental schemes. In many cases it is desirable to collect quantitative information ('hybridization signatures') rather than indications on 'positive' and 'negative' colonies. We present a practical system, based on an imaging plate analyser and a customized version of commercial software, that makes such quantification feasible, and define its performance in terms of reproducibility and linearity. The system is far superior to methods based on autoradiography and should be useful in many projects that involve the increasingly popular high density filter format.