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Any Goods not removed on or before the Removal Date shall be subject to storage charges that TPG may in its discretion impose.TPG does not pack, ship, store, crate or rig items. Buyers may either pick up Goods at the designated auction site personally or can contract with a third party agent to manage the removal process for them. TPG May from time to time, apply a loading charge. Before any removal, Buyer must provide TPG with a certificate of insurance from the rigger, which certificate must be in form, scope and content acceptable to TPG in TPG sole discretion. TPG shall have no responsibility to disconnect utilities to the sold asset, including electric, gas, waste and water lines. Buyer is solely responsible to properly remove and store in appropriate containers all fluids, oils, hazardous chemicals, etc., from machinery purchased. 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Abstract A prototype jig to attach a protein crystallization plate to a standard X-ray goniometer has been designed and constructed in partnership with an engineering firm. This allows a low-cost implementation of in situ diffraction using the available home-laboratory X-ray source. Keywords: in situ diffraction screening, crystallization plates, crystal quality, room-temperature diffraction 1.?IntroductionEarly testing of multiple hits from crystallization screening allows the selection and optimization of protein crystals based on diffraction quality rather than aesthetics. Even after optimization, it may be necessary to screen crystals in order to select those with the best resolution, lowest mosaicity or lowest anisotropy, or those with desirable cell and lattice parameters for further experiments. Compared to the usual practice of fishing a crystal from the mother liquor with a loop, and then cryoprotecting and flash-freezing it prior to X-ray diffraction testing, this technique offers two main advantages: (1) it avoids the damaging effects of crystal manipulation and (2) a room-temperature diffraction-quality baseline is established against which the effects of cryoprotectants and other soaking techniques can be compared. The in situ diffraction devices that are currently available tend to be highly automated both for synchrotron and for home X-ray sources. Oxford Diffraction (now Agilent) has developed the PX scanner ( ), which provides an integrated stand-alone solution for in situ diffraction including a sealed-tube X-ray source, X-ray optics and CCD detector for use in the home laboratory. The system allows the user to capture photographic images of crystals prior to automated X-ray screening, which is useful for correlating crystal morphology with diffraction quality. Diffraction images are processed using an algorithm that subtracts the scatter derived from the solvent and crystallization plate, resulting in an image that is easier to interpret. Lastly, as the system is fully integrated it is easily placed in any laboratory. Recent interest in in situ screening and data collection has prompted crystallization plate manufacturers to offer standard format plates with improved X-ray absorption characteristics ( e.g. CrystalQuickX; ). Better choices of plastic and careful positioning and design of the crystal wells have led to significant improvements in the overall intensity and angle of scattered X-rays. Innovative approaches to protein crystallization have led to a number of new devices on the market based on free interface diffusion or counter diffusion techniques. Hence, we devised a simple, manual, low-cost jig that allows attachment of a crystallization plate onto our Rigaku AFC-11 X-ray goniometer. 2.?Description of jigA crystallization plate is clipped into place on the plate holder, which is then slid into the base assembly previously mounted on the goniometer. A simple scale derived from the crystallization plate dimensions allows coarse positioning of the well of interest, while x, y, z micrometer adjustment screws allow accurate positioning of the crystal in the beam. Owing to the geometry of the Rigaku AFC-11 goniometer, it is only possible to access the bottom four rows of a plate held in landscape, so the jig allows the user to invert the plate and use a second scale to access the top four rows. Crystal centring exploits the existing Rigaku camera microscope and CameraMan software ( ). The cold light source on the Rigaku system was supplemented by a second light source to improve the image and help locate crystals in drops. Open in a separate window Figure 1 ( a ) Photograph of the jig standing on its base assembly. ( b ) Photograph of detached plate-holder assembly. ( c ) Photograph showing the pendulum beam stop assembly mounted on the Rigaku 944 CCD detector. The chassis, a, is made from plastic trunking, which accommodates a plastic pivot, b, from which the pendulum is hung. The beam stop is aligned by trial and error. A simple permanent pen mark on the acetate strip allows rapid and accurate repositioning after dismantling the setup. 3.?Typical in situ diffraction screening results using the jigCrystals were aligned manually in the X-ray beam using the Rigaku microscope camera and CameraMan display software.