RCT Volume¶

General Workflow:¶

Note: RCT Volume can be accessed directly from the Appion sidebar, or by clicking on the "Create RCT Volume" button displayed above class averages generated through 2D Alignment and Classification

1. Make sure that appropriate run names and directory trees are specified. Appion increments names automatically, but users are free to specify proprietary names and directories.
2. Enter a description of your run into the description box.
3. Select the aligned stack to use for titled particle parameters from the drop down menu. Note that stacks can be identified in this menu by clustering or feature analysis run ID. Also note that there will be no drop down menu for this option if the RCT Volume page was accessed directly from the clustering run output webpage, because these parameters are then already known.
4. Enter the class numbers to use for volume generation. To decide which classes to use, go look at your alignment/feature analysis/clustering output.
5. Select the TILTED particle stack to use for the reconstruction.
6. Specify a mask radius that is barely bigger than the radius of your molecule.
7. Click on "Rct Volume" to submit your job to the cluster. Alternatively, click on "Just Show Command" to obtain a command that can be pasted into a UNIX shell.
   

Notes, Comments, and Suggestions:¶

1. Specifying multiple classes will result in them being combined into a single RCT volume. Capability to launch multiple, separate RCT reconstructions at once coming soon!
2. The default parameters for median filter, low pass, and high pass can be changed. Number of particles uses only a subset of the data. The chimera settings only dictate the "look" of your output page. It does not alter the final outcome.

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