Supplementary Figure 1: Experimental reagents.
From: Dissection of DNA double-strand-break repair using novel single-molecule forceps
(A) Denaturing PAGE gel showing results of the oligo click reaction described in the Methods. Lane order is, left to right: DBCO oligo, Azido oligo, and DBCO and Azido oligos combined, for each of the two combinations of oligos used in the construct. (B) Schematic workflow for building construct from the Click-Oligos purified as in (A) (see Methods for details). (C) SDS-PAGE analysis of purified NHEJ components. 2 μg of each protein was loaded in each lane. “M” indicates protein ladder (kDa). (D) DNA ligation assays with ligase IV-XRCC4 and its catalytically dead mutant. 200 ng of linearized pHAT4 plasmid (double digested by XhoI and NcoI to give 4-bp overhangs) was incubated with 25 nM of ligase IV-XRCC4 and ligase IV(K273A)-XRCC4 (catalytically dead mutant). Agarose gel was stained with SYBR gold. See Methods for details.
