Summary
Overview of HT-X facilities
Facilities Technical Specification
 


 
Nanodrop dispensing for vapour diffusion experiments
 
Crystallography groups NKI Amsterdam
Dispensing Robot Plate type and experimental set up Well volume Drop volume (Protein + Well solution) Ammount of sample needed for 1 plate Ammount of sample needed for multiple plates Comments
Hydra II Dispenses crystallization solution (75 or 100 µl) in each well of a 96 well plate na (not applicable) na na na Crystallization solution is taken from 96 Deep-Well block
Mosquito Greiner low profile plate, 1 drop per crystallization well, sitting drop 75 µl 200 nl 28 µl: 8 rows x 3.5 µl/row 28 µl (first plate) + 20 µl (8 rows x 2.5 µl/row) for each additional plate Protein solutions are in excess; non-used protein solution can be recovered
Greiner plate, 3 drops per crystallization well, sitting drop: 1 protein solution 100 µl 200 nl 68 µl: 8 rows x (3.5 µl/row + 2.5 µl/row + 2.5 µl/row) 68 µl (first plate) + 60 µl (3 protein refills x 8 rows x 2.5 µl/row) for each additional plate The Mosquito has 2 pauses to refill protein solution in the strips
Greiner plate, 3 drops per crystallization well, sitting drop: 3 protein solutions 100 µl 200 nl 84 µl: 8 rows x 3.5 µl/row x 3 protein solutions 84 µl (first plate) + 60 µl (3 protein refills x 8 rows x 2.5 µl/row) for each additional plate The Mosquito has 2 pauses to refill protein solution in the strips
It takes less than a minute for the Mosquito to make drops in one 96 well plate
 
HTX lab EMBL Grenoble
Dispensing Robot Plate type and experimental set up Well volume Drop volume (Protein + Well solution) Ammount of sample needed for 1 plate Ammount of sample needed for multiple plates Comments
Tecan Te-Mo 3/5 96 channel pipetor Dispenses crystallization solution ( 88 µl) in each well of a 96 well plate na na na na Commercial Crystallization solutions are reformated from 10 ml tubes to deep well blocks (TECAN Genesis) and then to crystallization plates (TECAN TeMo)
Cartesian PixSys 4200 Plate Greiner Crystal Quick non-treated, sitting drop: 1, 2 or 3 samples 88 µl 200 nl 17 µl per sample 15 µl per sample per plate for two or more plates 8-well disposable strips are used to hold samples
Plate Greiner Crystal Quick non-treated and Polyolefin film. hanging drop: 1 or 2 samples 88 µl 200 nl 17 µl per sample 15 µl per sample per plate for two or more plates After a run of six plates ( 90 µl of sample ) 3 to 6 µl of sample can be recovered
Plate Greiner Crystal Quick non-treated, sitting drop: 3 sample/precipitant ratios 88 µl 200 nl 45 µl per sample 45 µl per sample per plate for two or more plates Up to 8 samples (3 crystallisation plates) can be processed in parallel
Experiments are performed under a humidity control chamber
In addition to the standard protocols described here a number of customized set ups are used
 
HT-X facility EMBL Hamburg
Dispensing Robot Plate type and experimental set up Well volume Drop volume (Protein + Well solution) Ammount of sample needed for 1 plate Ammount of sample needed for multiple plates Comments
Hydra II plusOne Reformat crystallization solution from DWP into any SBS format MTP into reservoir well (50 - 100 µL) and into crystallization well (200 - 1000 nL) Crystallization solutions are prepared into 96 DWP. Hydra reformats into MTP and adds protein. We could cut the protein loss to 1 µL
Greiner low profile, 1 drop per crystallization well, sitting drop 50 µL by choice, 300 nL most common at 200 nL: 22 µL; at 300 nL: 32 µL Makes no difference, as we wash between every step Many variations on standard protocol possible
Experiment exposure to air is 85s

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