Fluorescent Dyes and Dye Labelled Probes for Detection of Nucleic Acid Sequences in Biological Material

Abstract

The traditional method for the detection of nucleic acid sequences on membranes and in situ utilises radiolabelled compounds. These systems are still widely used in many applications, particularly where sensitivity and robustness are required1. However, in many situations there is less need for sensitivity and a more overriding need for resolution and speed of detection. To this end, effort has gone into the production and design of good, reliable, non-radioactive alternatives. Originally, these systems utilised hapten molecules, attached to nucleotides, to label the probes that were used to detect the presence of specific nucleic acid sequences. These hapten molecules were then detected through antibody conjugates. These too are now widely used in a number of applications2. In the field of chromosome in situ hybridization, detection through fluorescence labelled antibodies found great favour due to the ease of detection with modern microscopes and camera systems and, particularly with chromosome analysis, due to the low level of associated background fluorescence.