Measuring ligation activity of T4 DNA ligase on nicked DNA with switchSENSE^®

DNA ligases fulfill essential functions in DNA replication, DNA repair, and are standard tools in molecular biology labs. Here, we describe a switchSENSE^® assay for the rapid assessment of ligation activity with minimal sample consumption. The ligation activity of T4 DNA ligase is quantified in real-time by observing the ligation of a nick in one strand of a double stranded oligo. Complementary read-outs of dissociation behavior and melting temperature allow the reliable differentiation of ligated and non-ligated DNA oligos.

Using this assay, ligation activity or the influence of ligase inactivation, mutation or inhibition can be determined rapidly, accurately and in real-time for research, development and quality control.