Laboratory name
University of Virginia
Standard Virginia Crystalliation protcol:

1. After purification concentrate your sample to conc. 5-20 mg/ml using Amicon concentrators with proper MW membrane cut-off. It's better to prepare more concentrated samples because you can dilute it later
2. Aliquote your protein into samples of 10-50 ul and flash-freeze it in liquid nitrogen for future storage
3. Store your protein samples in -8- freezer in paper box
4. To set crystallization take fresh protein, or freezed one, then you need to thaw it on ice and then spin for 10 minutes, 3.000 rpm in 4C
5. Set screening crystallization using Mosquito
Standard screens used for basic screening are:
-Hampton Index

6. Pipette selected screens from tubes into 96 well block, 1 ml in each well. Tape it well for future storage, put name of the screen, your name and date
7. Using multichannel pipetor pipette 200ul of each condition into mosquito plate
8. Take 5ul strips and put 5 ul of protein into each well
9. Set crystallization with Mosquito
10. Tape plates with foil
11. Enter plate into Crystal Trak, print barcode, choose apropariate observation schedule and put plate into Minstrel
12. Observe plate
13. Enter plate into Xtaldb
14. If you get crystalls, optimize conditions
15. Use hanging/sitting drop Linbrio or Nextal plates (you can't observe Nextal plates via Minstrel)
16. Enter plate into Xtaldb, create pipetting guide
17. Set crystallization
18. Observe plate
19. Harvest crystalls

07/20/2009 Aleksandra Knapik