by Peter Nollert
August 31, 2010 06:55
How long does it take to set up a protein crystallization experiment?
It all depends on the equipment and the state of organization in your lab. Even with modest manual dexterity, setting up a single 96-well tray really shouldn't take more than 10 min. Allow 10 min of preparation time and you're at only 20 min. In fact, without the use of robotics it is possible to process a 40 uL protein sample and prepare a 96-well protein crystallization trial within less than 20 minutes, all things considered. Multiply that by 2 or 4 if you aim to increase coverage of crystallization phase space. Here's a rough schedule for preparing a single tray consisting of 96 x 0.8 uL sitting drop protein crystallization experiments using the vapor diffusion method :
| Activity |
Time |
|
Dispensing the well-solutions from a 96 well block (i.e. a Wizard III & IV) into a 96-well sitting drop plate (i.e. Clompact Jr. plates) using a 12 channel pipettor = 8 transfer steps
|
2-3 min |
|
Dispensing 96 x 0.4 uL of protein solution into the crystallization chamber using multiple volume pipettor (a P20 takes only 3 refills)
|
2-3 min |
|
Transferring 0.4 uL of well solution to the crystallization chamber using a 12 channel pipettor = 8 transfer steps
|
2-3 min |
|
Sealing of tray by attaching clear adhesive tape
|
< 1 min |
|
Preparation time: setting up work space, getting tip boxes ready, unpacking source block and crystallization tray, collection of liquids in the source block with a short spin, removal and application of the cap mat.
|
ca. 10 min (depends on organization in the lab) |
Is setting up a single crystallization tray taking you more than 20 min? Maybe it's worthwhile getting those multichannel pipettors and repeating dispensers, or switching from slow hanging drops to fast sitting drops.
Or just clean up the lab and get those utensils that you already have, better organized ? ;)
Cheers, Peter