Overview
Synthetic oligonucleotides lacking phosphate groups at their 5' termini are easily radiolabeled by transfer of the {gamma}-32P from [{gamma}-32P]ATP in a reaction catalyzed by bacteriophage T4 polynucleotide kinase. The reaction described below is designed to label 10 pmoles of an oligonucleotide to high specific activity. Labeling of different amounts of oligonucleotide can easily be achieved by increasing or decreasing the size of the reaction while keeping the concentrations of all components constant. When the reaction is carried out efficiently, >50% of the oligonucleotide molecules in the reaction become radiolabeled. By Joseph Sambrook and David W. Russell.
Details
Details of this protocol, Phosphorylating The 5' Termini Of Oligonucleotides (Subscription Required), are located on a web site other than Biocompare Protocols.