Towards rationalization of crystallization screening for small- to medium-sized academic laboratories: the PACT/JCSG+ strategy.
A crystallization screening process is presented that was developed for a small academic laboratory. Its underlying concept is to combine sparse-matrix screening with systematic screening in a minimum number of crystallization conditions. The sparse-matrix screen is the cherry-picked combination of conditions from the Joint Center for Structural Genomics (JCSG) extended using conditions from other screens. Its aim is to maximize the coverage of crystallization parameter space with no redundancy. The systematic screen, a pH-, anion- and cation-testing (PACT) screen, aims to decouple the components of each condition and to provide information about the protein, even in the absence of crystals, rather than cover a wide crystallization space. This screening strategy is combined with nanolitre-volume dispensing hardware and a small but practical experiment-tracking system. The screens have been tested both at the NKI and in other laboratories and it is concluded that they provide a useful minimal screening strategy.
Newman
Janet
J
Egan
David
D
Walter
Thomas S
TS
Meged
Ran
R
Berry
Ian
I
Ben Jelloul
Marouane
M
Sussman
Joel L
JL
Stuart
David I
DI
Perrakis
Anastassis
A
D000465:Algorithms; D000838:Anions; D001331:Automation; D002412:Cations; D002621:Chemistry / Q000379:methods*; D056246:Chemistry, Analytic / Q000379:methods*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000295:instrumentation* / Q000379:methods*; D030562:Databases, Protein; D006863:Hydrogen-Ion Concentration; D007753:Laboratories; D011506:Proteins; D012107:Research Design; D012984:Software; D014495:Universities
D000838:Anions; D002412:Cations; D011506:Proteins