PMID- 15930615
OWN - NLM
STAT- MEDLINE
DCOM- 20050818
LR  - 20211109
IS  - 0907-4449 (Print)
IS  - 1399-0047 (Electronic)
IS  - 0907-4449 (Linking)
VI  - 61
IP  - Pt 6
DP  - 2005 Jun
TI  - A procedure for setting up high-throughput nanolitre crystallization experiments. 
      Crystallization workflow for initial screening, automated storage, imaging and 
      optimization.
PG  - 651-7
AB  - Crystallization trials at the Division of Structural Biology in Oxford are now 
      almost exclusively carried out using a high-throughput workflow implemented in 
      the Oxford Protein Production Facility. Initial crystallization screening is 
      based on nanolitre-scale sitting-drop vapour-diffusion experiments (typically 100 
      nl of protein plus 100 nl of reservoir solution per droplet) which use standard 
      crystallization screening kits and 96-well crystallization plates. For 294 K 
      crystallization trials the barcoded crystallization plates are entered into an 
      automated storage system with a fully integrated imaging system. These plates are 
      imaged in accordance with a pre-programmed schedule and the resulting digital 
      data for each droplet are harvested into a laboratory information-management 
      system (LIMS), scored by crystal recognition software and displayed for user 
      analysis via a web-based interface. Currently, storage for trials at 277 K is not 
      automated and for imaging the crystallization plates are fed by hand into an 
      imaging system from which the data enter the LIMS. The workflow includes two 
      procedures for nanolitre-scale optimization of crystallization conditions: (i) a 
      protocol for variation of pH, reservoir dilution and protein:reservoir ratio and 
      (ii) an additive screen. Experience based on 592 crystallization projects is 
      reported.
FAU - Walter, Thomas S
AU  - Walter TS
AD  - Oxford Protein Production Facility, Division of Structural Biology, Henry 
      Wellcome Building for Genomic Medicine, Roosevelt Drive, Headington, Oxford OX3 
      7BN, England.
FAU - Diprose, Jonathan M
AU  - Diprose JM
FAU - Mayo, Chris J
AU  - Mayo CJ
FAU - Siebold, Christian
AU  - Siebold C
FAU - Pickford, Mike G
AU  - Pickford MG
FAU - Carter, Lester
AU  - Carter L
FAU - Sutton, Geoff C
AU  - Sutton GC
FAU - Berrow, Nick S
AU  - Berrow NS
FAU - Brown, James
AU  - Brown J
FAU - Berry, Ian M
AU  - Berry IM
FAU - Stewart-Jones, Guillaume B E
AU  - Stewart-Jones GB
FAU - Grimes, Jonathan M
AU  - Grimes JM
FAU - Stammers, David K
AU  - Stammers DK
FAU - Esnouf, Robert M
AU  - Esnouf RM
FAU - Jones, E Yvonne
AU  - Jones EY
FAU - Owens, Ray J
AU  - Owens RJ
FAU - Stuart, David I
AU  - Stuart DI
FAU - Harlos, Karl
AU  - Harlos K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20050526
PL  - United States
TA  - Acta Crystallogr D Biol Crystallogr
JT  - Acta crystallographica. Section D, Biological crystallography
JID - 9305878
RN  - 0 (Proteins)
SB  - IM
MH  - Animals
MH  - Automation/instrumentation/methods
MH  - Crystallography, X-Ray/instrumentation/*methods
MH  - Humans
MH  - Nanotechnology/instrumentation/*methods
MH  - Proteins/*chemistry
PMC - PMC7159505
EDAT- 2005/06/03 09:00
MHDA- 2005/08/19 09:00
PMCR- 2020/04/15
CRDT- 2005/06/03 09:00
PHST- 2005/06/03 09:00 [pubmed]
PHST- 2005/08/19 09:00 [medline]
PHST- 2005/06/03 09:00 [entrez]
PHST- 2020/04/15 00:00 [pmc-release]
AID - S0907444905007808 [pii]
AID - AYDIC5008 [pii]
AID - 10.1107/S0907444905007808 [doi]
PST - ppublish
SO  - Acta Crystallogr D Biol Crystallogr. 2005 Jun;61(Pt 6):651-7. doi: 
      10.1107/S0907444905007808. Epub 2005 May 26.