Evaluation of non-radioactive labelling and detection of deoxyribonucleic acids. Part Two: Colorigenic methods and comparison with chemiluminescent methods.
The diagnosis of genetic infections and cancerous diseases is carried out more and more often at a molecular level using Southern's technique which is based on the use of 32P-labelled DNA. In order to circumvent the risks and rapid decrease in radioactivity associated with these latter techniques, new colorigenic methods have been developed. In this work, we describe the use of dTTP analogues (digoxigenin-dUTP and biotin-dUTP) for the labelling of probes and detection of target DNA. Using digoxigenin-11-dUTP, 0.1 aM of a 561 bp target DNA was detected by using a modified Southern procedure. The reliability and the high sensitivity of such methods make them a good tool for DNA investigation in research as well as in testing laboratories.
D000230:Adenocarcinoma / Q000737:chemistry; D000818:Animals; D015139:Blotting, Southern; D003101:Collodion; D003124:Colorimetry; D004247:DNA / Q000032:analysis*; D004273:DNA, Neoplasm / Q000032:analysis; D004076:Digoxigenin; D005784:Gene Amplification; D008163:Luminescent Measurements; D008325:Mammary Neoplasms, Experimental / Q000737:chemistry; D008567:Membranes, Artificial; D051379:Mice; D012680:Sensitivity and Specificity
D004273:DNA, Neoplasm; D008567:Membranes, Artificial; D003101:Collodion; D004247:DNA; D004076:Digoxigenin