Fig. 7: Preparation of dual labeled dsDNA and T7 RNA polymerase-promoter cross-linking.
From: Programmable site-selective labeling of oligonucleotides based on carbene catalysis
a DNA-protein cross-linking of the probe and DNA-binding protein in the absence (b and c) and in the presence of E. coli lysate (d and e). b Oligonucleotide probes employed for c. c SDS–PAGE analysis of the cross-linking reaction visualized by silver and SYBR gold stains. Conditions: dsDNA (20 µM), T7 RNA polymerase (T7 RNAP) (6.8 µM), HEPES (40 mM), pH 7.4, Mg(OAc)2 (5 mM), NaCNBH3 (100 mM), 37 °C, 12 h. d Synthetic scheme for dual G-bulge probe 22a employed for e. e SDS–PAGE analysis of DNA-protein cross-linking in E. coli lysate. Conditions: dsDNA (20 µM), T7 RNAP (3.4 µM, for 1x), HEPES (40 mM), pH 7.4, Mg(OAc)2 (5 mM), NaCNBH3 (100 mM), 30% E. coli lysate, 37 °C, 12 h. †unmodified oligo was used. L: protein standard ladder. Source data are provided as a Source Data file.
