5 fam labeled single stranded dna ssdna oligonucleotides

The gel shift assay was adapted from 5. 5′‐FAM‐labeled single‐stranded DNA (ssDNA) oligonucleotides were purchased from Integrated DNA Technologies (Coralville, IA, USA). Oligonucleotides (10 μm) were folded in folding buffer (20 mm Tris pH 7.5, 100 mm KCl, 1 mm EDTA) by heating at 95 °C for 5 min, then allowed to cool passively to room temperature. Folded DNA (0.6 μm) was mixed with recombinant NME2 protein in reaction buffer (50 mm Tris pH 8.0, 100 mm KCl, 1 mm TCEP, 50 μg·mL⁻¹ BSA) at room temperature for 30 min, followed by electrophoresis on 5% native polyacrylamide TBE gel in 0.5× TBE at 70 V for 1 h and analyzed using Typhoon FLA7000 scanner (GE Healthcare). For sequences of oligonucleotides used, see Fig. S3.