Specific chemical labeling of DNA fragments.
We describe a simple method for specific chemical labeling of DNA fragments at their 3'-termini. The procedure includes enzymatic addition of 4-thiouridine, followed by reaction in mild non-denaturing conditions with the highly reactive alpha-haloacetamido derivatives of several chemical labels. The attached reporter molecule can be removed by extended treatment with beta-mercaptoethanol. Among the potential applications of this labeling method is the study of specific protein-DNA interactions in solution.
D055598:Chemical Phenomena; D002621:Chemistry; D004247:DNA; D004254:DNA Nucleotidyltransferases; D004801:Eosine Yellowish-(YS) / Q000031:analogs & derivatives*; D005452:Fluoresceins; D007460:Iodoacetamide / Q000031:analogs & derivatives; D008722:Methods; D009282:Naphthalenesulfonates; D013891:Thiouridine
D005452:Fluoresceins; D009282:Naphthalenesulfonates; D013891:Thiouridine; D004247:DNA; D004254:DNA Nucleotidyltransferases; D004801:Eosine Yellowish-(YS); D007460:Iodoacetamide