Production and purification of tagmentation-ready Tn5. (A) Illustration of the Tn5–intein fusion construct. (B) 10% SDS-PAGE run with crude lysate (Lys), supernatant (Sup), flow-through (FT), wash (W), and DTT eluted fractions (DTT) from the chitin column. The expected sizes of the Tn5–intein fusion protein (Mxe-Tn5) and cleaved full-length Tn5 (Tn5) are indicated on the right; marker molecular weights in kDa, to the left. (C) Agarose gel (1% Type LE, in TAE buffer) demonstrating tagmentation of 50 or 100 ng high MW calf thymus DNA with 1 μL of Tn5 prepared by annealing in solution (Free), on chitin columns (Col), or with no Tn5 as a negative control (C). On-column annealing removed excess oligonucleotides present after solution annealing.
