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Crystal Screening and Data Collection Activities at SDC. Jessica Chiu Joint Center for Structural Genomics Stanford Synchrotron Radiation Laboratory JCSG 5 th annual meeting La Jolla, CA, April 6-7,2006. Beamtime provision during SSRL run time. Weekly access to two of the 6 PX beamlines

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Crystal screening and data collection activities at sdc

Crystal Screening and Data Collection Activities at SDC

Jessica Chiu

Joint Center for Structural Genomics

Stanford Synchrotron Radiation Laboratory

JCSG 5th annual meeting

La Jolla, CA, April 6-7,2006


Beamtime provision during SSRL run time

Weekly access to two of the 6 PX beamlines

All beamlines with automated sample mounting

4 beamlines capable of MAD experiments

Primary

Access

Crystal screening

Screen ~15,000 crystals per year

Data collection

Collect ~400

Datasets per year

Storage ring upgraded in 2003/4 (SPEAR-3)

Plan to increase from 100mA to 500mA current


Beamtime provision during SSRL shutdown periods

2-3 days /

quarter

GM/CA CAT

Secondary

Access

Required during SSRL shutdownperiods (2-3 months per year).

Crystal screening continues at SSRLand pre-screened crystals are taken

to other synchrotron facilities.

Data collection

Collect ~400datasets per year

Crystal screening

Screen ~15000 crystals per year

Fully automated 24/7

crystal screening facility.

Based around X-ray microsource generator.

Temporarily installed in SSRL BL9-2 hutch each shutdown.

Will develop into permanent offline screening facility in 2006

~3 days /

month


Crystal Screening statistics by year

5127

1884

6807

4253*

6262

1530

235

# crystals screened

~21000 crystals screened in total

*The number is lower than expected due to SSRL Spear III

upgrade and safety stand-down at SLAC.


Crystal Screening: where and how?

56%

30%

3%



10%

1%

1%



Screened crystals used for data collection

Average screen resolution = 2.46Å

Average collect resolution = 2.31 Å

For 430 crystals/datasets

Average screen resolution = 3.67Å

Average collect resolution = 2.30 Å

For 193 crystals/datasets




Crystal screening to test different crystallization strategies

*Structures solved are in bold.


Database tools for managing crystal screening strategies

end date

Crystal Screening Activity Report

start date

list for every target

  • Cassette/Dewar Tracking Tool

  • cassette/dewar inventory

  • synchronized with beamline report

  • shipment tracking and automated

  • email notification


Summary strategies

  • Crystal shipments are well coordinated with beam time schedule.

  • Various database tools were developed to help management.

  • With current staffing (1 100% RA, 1 30% scientist), we are capable of processing 300 crystals per week.

  • We have been working closely with Crystallomics core on managing solved target list. The list is updated weekly with targets “pause”, “stop” or “send”. This increase efficiency in both CC and SDC.


Future improvements
Future improvements strategies

  • Actively manage marginal targets

    • Provide feedback on salvage pathways (promising crystallization conditions, protein variation etc) to guide ongoing crystallization experiments

  • Target screening history report

    • Provide a concise, but informative report for the screening results on individual targets.

  • Fine screening

    • Human effort to focus on the individual problematic targets.


SDC strategies

Keith Hodgson

Ashley Deacon

Silvya Oommachen

Qinging Xu

Mitchell Miller

Herbert Axelrod

Christopher Rife

Kevin Jin

Henry van den Bedem

Abhinav Kumar

Jonathan Caruthers

Chloe Zubieta

CC

Scott lesley

Mark Kunth

Dennis Carlton

Eileen Ambing

Linda Okach

Daniel McMullan

Heath Klock

Michael DiDonato

Tom Clayton

BIC

Adam Godzik

Slawek Grzechnik

Andrew Morse

Lian Duan

Bill West

AC

Ian Wilson

Marc-Andre Elsliger

Jason Kay

SSRL

Jinhu Song Craeme Card

Scott McPhillips Clyde Smith

Mathews Irimpan Penjit Moorhead

Ana Gonzalez Thomas Eriksson

Aina Cohen

Michael Solits

Lisa Dunn

NIH Protein Structure Initiative Grant P50 GM62411


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