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MICROFLUIDIC DEVICES AND METHODS FOR PROTEINS CRYSTALLIZATION AND IN SITU X-RAY DIFFRACTION
MICROFLUIDIC DEVICES AND METHODS FOR PROTEINS CRYSTALLIZATION AND IN SITU X-RAY DIFFRACTION
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机译:蛋白质结晶和原位X射线衍射的微流体装置和方法
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摘要
The present disclosure is directed generally to devices and methods with the purpose of interfacing micro fluidic devices with dispensing and fluid handling systems to achieve the rapid identification of protein crystallization conditions. The device described herein is fabricated with the use of a cyclic olefin homopolymer-based creating microfluidics system adaptable for protein crystallization and in situ X-ray diffraction. Connectivity between chambers is controlled by valves that allow specified volumes of liquid to be transferred from one chamber to another. The microfluidic system is useful to established microbatch, vapor diffusion and free interface diffusion protocols for protein crystallization and to obtain crystals for a number of proteins, including chicken lysozyme, bovine trypsin, a human p53 protein containing both the DNA binding and oligomerization domains bound to DNA and a functionally important domain of Arabidopsis Morpheus' Molecule 1 (MOMl). For X-ray diffraction analysis, either the microfluidic devices were opened to allow mounting of the crystals on loops or the crystals were exposed to X-rays in situ. Thus, cyclic olefin homopolymer-based microfluidics systems are useful to further automate protein crystallization and structural genomics efforts.
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