Monitoring Incorporation of Fluorescein into Nucleic Acid Probes Using a Rapid Labeling Assay

The efficiency of incorporation of fluorescein-11-dUTP with both random prime labeling of long probes (Chapter 8 ) and 3′-tailing of oligonucleotide probes (Chapter 11 ) can be estimated using the following rapid labeling assay. The basic protocol shown is for use with random prime labeled long probes, but modifications to the protocol for use with oligonucleotides are detailed in the notes. If sample identification is required, a pencil should be used to mark the membrane since some inks can interfere with the assay.