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Title: Automated diffraction analysis and spot searching forhigh-throughput crystal screening

Abstract

No abstract prepared.

Authors:
; ; ; ;
Publication Date:
Research Org.:
COLLABORATION - Stanford Synchrotron RadiationLaboratory
OSTI Identifier:
927186
Report Number(s):
LBNL-60815
Journal ID: ISSN 0021-8898; JACGAR; R&D Project: 864D6D; TRN: US200810%%160
DOE Contract Number:
DE-AC02-05CH11231
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Applied Crystallography; Journal Volume: 39; Journal Issue: 1; Related Information: Journal Publication Date: February2006
Country of Publication:
United States
Language:
English
Subject:
59; 47; X-RAY DIFFRACTION; AUTOMATION; CRYSTAL STRUCTURE; crystallography X-ray diffraction

Citation Formats

Zhang, Zepu, Sauter, Nicholas K., van den Bedem, Henry, Snell,Gyorgy, and Deacon, Ashley M.. Automated diffraction analysis and spot searching forhigh-throughput crystal screening. United States: N. p., 2005. Web.
Zhang, Zepu, Sauter, Nicholas K., van den Bedem, Henry, Snell,Gyorgy, & Deacon, Ashley M.. Automated diffraction analysis and spot searching forhigh-throughput crystal screening. United States.
Zhang, Zepu, Sauter, Nicholas K., van den Bedem, Henry, Snell,Gyorgy, and Deacon, Ashley M.. Thu . "Automated diffraction analysis and spot searching forhigh-throughput crystal screening". United States. doi:.
@article{osti_927186,
title = {Automated diffraction analysis and spot searching forhigh-throughput crystal screening},
author = {Zhang, Zepu and Sauter, Nicholas K. and van den Bedem, Henry and Snell,Gyorgy and Deacon, Ashley M.},
abstractNote = {No abstract prepared.},
doi = {},
journal = {Journal of Applied Crystallography},
number = 1,
volume = 39,
place = {United States},
year = {Thu Dec 08 00:00:00 EST 2005},
month = {Thu Dec 08 00:00:00 EST 2005}
}
  • No abstract prepared.
  • The macromolecular crystallography experiment lends itself perfectly to high-throughput technologies. The initial steps including the expression, purification, and crystallization of protein crystals, along with some of the later steps involving data processing and structure determination have all been automated to the point where some of the last remaining bottlenecks in the process have been crystal mounting, crystal screening, and data collection. At the Stanford Synchrotron Radiation Laboratory, a National User Facility that provides extremely brilliant X-ray photon beams for use in materials science, environmental science, and structural biology research, the incorporation of advanced robotics has enabled crystals to be screenedmore » in a true high-throughput fashion, thus dramatically accelerating the final steps. Up to 288 frozen crystals can be mounted by the beamline robot (the Stanford Auto-Mounting System) and screened for diffraction quality in a matter of hours without intervention. The best quality crystals can then be remounted for the collection of complete X-ray diffraction data sets. Furthermore, the entire screening and data collection experiment can be controlled from the experimenter's home laboratory by means of advanced software tools that enable network-based control of the highly automated beamlines.« less
  • We have developed an automated purification method for terminator sequencing products based on a magnetic bead technology. This 384-well protocol generates labeled DNA fragments that are essentially free of contaminates for less than $0.005 per reaction. In comparison to laborious ethanol precipitation protocols, this method increases the phred20 read length by forty bases with various DNA templates such as PCR fragments, Plasmids, Cosmids and RCA products. Our method eliminates centrifugation and is compatible with both the MegaBACE 1000 and ABIPrism 3700 capillary instruments. As of September 2001, this method has produced over 1.6 million samples with 93 percent averaging 620more » phred20 bases as part of Joint Genome Institutes Production Process.« less
  • A robotic system for auto-mounting crystals from liquid nitrogen is now operational on SSRL beamlines (Cohen et al., J. Appl. Cryst. (2002). 35, 720-726). The system uses a small industrial 4-axis robot with a custom built actuator. Once mounted, automated alignment of the sample loop to the X-ray beam readies the crystal for data collection. After data collection, samples are returned to the cassette. The beamline Dewar accommodates three compact sample cassettes (holding up to 96 samples each). During the past 4 months, the system on beamline 11-1 has been used to screen over 1000 crystals. The system has reducedmore » both screening time and manpower. Integration of the hardware components is accomplished in the Distributed Control System architecture developed at SSRL (McPhillips et al., J. Synchrotron Rad. (2002) 9, 401-406). A crystal-screening interface has been implemented in Blu-Ice. Sample details can be uploaded from an Excel spreadsheet. The JCSG generates these spreadsheets automatically from their tracking database using standard database tools (http://www.jcsg.org). New diffraction image analysis tools are being employed to aid in extracting results. Automation also permits tele-presence. For example, samples have been changed during the night without leaving home and scientists have screened crystals 1600 miles from the beamline. The system developed on beamline 11-1 has been replicated onto 1-5, 9-1, 9-2, and 11-3 and is used by both general users and the JCSG.« less