Characterization of Action and Efficiency of Tn5 Transposase Through Comparative qPCR

We describe initial functional profiling of an in-house cloned Tn5 transposase in the transposition based sequencing library preparation reaction termed tagmentation.  We provide the basis for a quantitative procedure to compare efficiency of different Tn5 enzymes in the tagmentation reaction. Fragment size distributions after tagmentation by our transposase varied greatly from those previously described utilizing Illumina’s Nextera derivative, suggesting differences in enzyme activity. A comparative qPCR assay of tagmentation efficiency was unable to resolve these differences present within our tagmentation libraries after reaction attenuation, however a simulation of differential fragment length, created by library size selection, was resolvable by this method. Together, our findings highlight the need of a quantitative transposase activity assay, their feasibility, and the difficulty of providing such assays in a cost-effective high-throughput manner.