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    Nat Protoc. 2008;3(12):1941-74.

    SPIDER image processing for single-particle reconstruction of biological macromolecules from electron micrographs.

    Shaikh TR, Gao H, Baxter WT, Asturias FJ, Boisset N, Leith A, Frank J.

    Source

    Wadsworth Center, Empire State Plaza, Albany, New York 12201-0509, USA.

    Abstract

    This protocol describes the reconstruction of biological molecules from the electron micrographs of single particles. Computation here is performed using the image-processing software SPIDER and can be managed using a graphical user interface, termed the SPIDER Reconstruction Engine. Two approaches are described to obtain an initial reconstruction: random-conical tilt and common lines. Once an existing model is available, reference-based alignment can be used, a procedure that can be iterated. Also described is supervised classification, a method to look for homogeneous subsets when multiple known conformations of the molecule may coexist.

    PMID:
    19180078
    [PubMed - indexed for MEDLINE]
    PMCID:
    PMC2737740
    Free PMC Article

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