Oryx 6 dispensing routines
Home Up Oryx Spec Picture of Oryx Oryx 6 dispensing routines Microbatch Crystallization Getting Started with Oryx MCC Spec

 

Summary of Components and dispensing routines
for Oryx 6

 

Hardware Components of Oryx 6

 

         Microtip

         Disposable tip

         Large table able to hold 3 Cryschem plates or the equivalent

         5 small motorized syringes (each holding up to 117 μl)

         1 large motorized syringe (10 ml)

         Ground glass syringes to refill the motorized syringes with water.

         Disposable syringes to flush the microtip and to load it with stock solutions

         MCC Controller (with 12 motor outputs).

 

Dispensing Routines for Oryx 6

 

a. Microbatch screening

 

1.   A 2-bore microtip is used.  (A 1ml disposable tip is used to add oil.)

2.   Just enough protein for the experiment (normally 10 to 80 μl) is loaded into the first bore at the start of the experiment, by sucking it into the tip.

3.   The tip goes to the first cocktail.  *0.1-2.0 μl is picked up into the second bore.

4.   The tip goes to the first target well. 0.1-2.0 μl of protein is dispensed at the same time as the 0.1-2.0 μl of cocktail.

5.   The experiment is stirred by moving the microtip in a 4-leaf clover pattern.

6.   12 μl of oil is dispensed onto the experiment to prevent evaporation

7.   The microtip is rinsed.

8.   Steps 3 8 are repeated, for 48 to 384 cocktails.

 

* Oryx 0.1-2.0 μl; IMPAX 0.3-2.0 μl

 

b. Microbatch optimization

 

1.   A 5-bore microtip is used. (A 1ml disposable tip is used to add oil.)

2.   The user is instructed to prime channel 2 with precipitant, channel 3 with buffer, and channel 4 with additive (or buffer at a different pH for pH gradients).  Channel 5 always contains water.

3.   Just enough protein for the experiment (normally 10 to 70 μl) is loaded into the first channel.

4.   Droplets are rapidly dispensed to the wells (all solutions are dispensed at the same time).

5.   After each droplet is dispensed, oil is dispensed onto it.

 

c. Sitting drops for screening and optimization.

 

1.   A 2-bore microtip is used.

2.   Just enough protein for the experiment (normally 10 to 110 μl) is loaded into the first bore at the start of the experiment, by sucking it into the tip.

3.   The tip goes to the first reservoir.  Around 1 μl is picked up into the second bore.

4.   The tip goes to the first target well. Around 1 μl of protein is dispensed at the same time as the 1 μl of cocktail.

5.   The experiment is stirred by moving the microtip in a 4-leaf clover pattern.

6.   The microtip is rinsed in the next reservoir (this is done with an offset to prevent contamination of the material that is transferred to the droplet).

7.   Steps 3 6 are repeated for up to 96 wells.

 

Filling reservoirs for Sitting Drop screens

1.   A 1ml disposable tip is used.

2.   Solutions are transferred from a mother plate to a daughter plate (e.g. a sitting drop plate).

3.   The tip is rinsed in water or buffer after each transfer.
 

                                                 2004 Douglas Instruments Ltd