Sybr green 1 stain

All chemicals were sourced from Sigma Chemical Co. (St. Louis, MO) unless stated otherwise. Sodium ascorbate was from Acros Organics (Geel, Belgium) while hydrogen peroxide (30%) was purchased from Thermo Fisher Scientific (Waltham, MA). Electron microscopy grade paraformaldehyde solution (16%) was obtained from Electron Microscopy Sciences (Hatfield, PA). NdeI was purchased from New England Biolabs (Beverly, MA). SYBR green I stain and Alexa fluor azide reagents (488 and 546, US Patents US20080050731 A1) were from Life Technologies (Carlsbad, CA). Vectashield mounting medium was from Vector Laboratories (Burlingame, CA). Propidium iodide was obtained from Roche Life Sciences (Indianapolis, IA). Micro Bio-Spin 6 chromatography columns were purchased from Bio-Rad (Hercules, CA). T2AA ((S)-4-(4-(2-Amino-3-hydroxypropyl)-2,6-diiodophenoxy)phenol hydrochloride) was prepared as previously described [17 (link)]. Olaparib was obtained from Selleck Chemicals (Houston, TX). A QIAquick PCR Purification Kit was from Qiagen (Valencia, CA). A pEGFP-PCNA plasmid was originally obtained from Daniel Gerlich [18 (link)] via Addgene (Cambridge, MA).

Chemical libraries for HTS were acquired from various sources. An additional batch of compound 1 was purchased from ChemBridge Corp. (San Diego, CA) for revalidation. All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO) unless stated otherwise. Restriction DNA endonucleases and DNA ligases were purchased from New England Biolabs (Beverly, MA). SYBR green I stain was purchased from Life Technologies (Carlsbad, CA). The AlphaScreen assay kit was purchased from PerkinElmer (Waltham, MA). Taq DNA polymerases for PCR, the PCR purification kit, and the Plasmid Miniprep kit were purchased from Qiagen (Valencia, CA). COS7 and HeLa cells were obtained from American Type Culture Collection (Manassas, VA) and cultured in Dulbecco’s Modified Eagle Medium containing 10% FBS. All cells were maintained in an incubator at 37°C in a humidified atmosphere of 5% carbon dioxide.