Semi-automated microseeding of nanolitre crystallization experiments.
A simple semi-automated microseeding procedure for nanolitre crystallization experiments is described. Firstly, a microseed stock solution is made from microcrystals using a Teflon bead. A dilution series of this microseed stock is then prepared and dispensed as 100 nl droplets into 96-well crystallization plates, facilitating the incorporation of seeding into high-throughput crystallization pipelines. This basic microseeding procedure has been modified to include additive-screening and cross-seeding methods. Five examples in which these techniques have been used successfully are described.
Walter
Thomas S
TS
Oxford Protein Production Facility and Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, England.
Mancini
Erika J
EJ
Kadlec
Jan
J
Graham
Stephen C
SC
Assenberg
René
R
Ren
Jingshan
J
Sainsbury
Sarah
S
Owens
Raymond J
RJ
Stuart
David I
DI
Grimes
Jonathan M
JM
Harlos
Karl
K
D001331:Automation; D001426:Bacterial Proteins / Q000737:chemistry*; D018360:Crystallography, X-Ray / Q000379:methods; D029968:Escherichia coli Proteins / Q000737:chemistry; D023281:Genomics; D008958:Models, Molecular; D008968:Molecular Conformation; D036103:Nanotechnology / Q000379:methods*; D040901:Proteomics; D014764:Viral Proteins / Q000737:chemistry*
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D014764:Viral Proteins