The assembly of the Tn5 and T7-Tn5 transposases were performed as described (Picelli et al. 2014 (link)). Briefly, oligonucleotides (T7-Tn5ME, Tn5MErev, Tn5ME-A, Tn5ME-B) were resuspended in water to a final concentration of 100 µM each. Equimolar amounts of Tn5MErev/Tn5ME-T7, Tn5MErev/Tn5ME-A, and Tn5MErev/Tn5ME-B were mixed in separate 200-µL PCR tubes. These oligos mixtures were denatured on a thermocycler for 5 min at 95°C and cooled down slowly on the thermocycler by turning off the thermocycler. The T7-Tn5 transposase was assembled with the following components: 0.25 vol Tn5MErev/Tn5ME-T7 (final concentration of each double-strand oligo is now 50 µM each), 0.4 vol glycerol (100% solution), 0.12 vol 2× dialysis buffer (100 mM HEPES-KOH at pH 7.2, 0.2 M NaCl [Invitrogen AM9759], 0.2 mM EDTA [Invitrogen AM9290G], 2 mM DTT, 0.2% Triton X-100 [Sigma-Aldrich T8787] 20% glycerol [Sigma-Aldrich G9012-500]), 0.1 vol SL-Tn5 (50 µM), 0.13 vol water. The reagents were mixed thoroughly but gently, and the solution was left on the bench for 1 h at room temperature to allow annealing of oligos to Tn5. The Tn5 transposase was assembly with same procedure as T7-Tn5 transposase but with following oligos: 0.25 vol Tn5MErev/Tn5ME-A and 0.25 vol Tn5MErev/Tn5ME-B.