Author: Tan, Jinzhi; Vonrhein, Clemens; Smart, Oliver S.; Bricogne, Gerard; Bollati, Michela; Kusov, Yuri; Hansen, Guido; Mesters, Jeroen R.; Schmidt, Christian L.; Hilgenfeld, Rolf
Title: The SARS-Unique Domain (SUD) of SARS Coronavirus Contains Two Macrodomains That Bind G-Quadruplexes Document date: 2009_5_15
ID: 1aqt65cc_35
Snippet: Crystallization SUD core displayed .95% purity in SDS-PAGE, and monodispersity in Dynamic Light-Scattering. Initial crystallization screening was performed using the sitting-drop vapor-diffusion method in 96-well Intelli-Plates (Dunn Laboratories). Several commercial kits (Sigma, Jena Bioscience) were used for the screening. The protein concentration was 6 mg/ml. Using a Phoenix robotic system (Art Robbins), drops were made of 260 nl protein and .....
Document: Crystallization SUD core displayed .95% purity in SDS-PAGE, and monodispersity in Dynamic Light-Scattering. Initial crystallization screening was performed using the sitting-drop vapor-diffusion method in 96-well Intelli-Plates (Dunn Laboratories). Several commercial kits (Sigma, Jena Bioscience) were used for the screening. The protein concentration was 6 mg/ml. Using a Phoenix robotic system (Art Robbins), drops were made of 260 nl protein and 260 nl precipitant solution. The optimized crystallization condition consisted of 20% polyethylene glycol monomethyl ether 5000 and 0.2 M ammonium sulfate in 0.1 M morpholinoethane sulfonic acid (pH 6.5). Plate-like crystals grew in 3-5 days, to maximum dimensions of 0.0260.0260.01 mm 3 .
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