Reproducing HWI crystallization screening hits
We recommend a Greiner 72-well microbatch plate, treated hydrophilic (Hampton Research HR3‐121).
Set up 5 replications each of experiments (1) through (3) by following the protocol described below.
- 1μL protein + 2μL cocktail
- 1μL protein + 1μL cocktail
- 2μL protein + 1μL cocktail
Step 1. Add the cocktail drop to each of the five replicate experiment wells in a row of the plate. You can use a microscope to watch the drop deliveries to each set up if desired.
Step 2. Add the protein drop (do not mix) by touching the pipette tip to the cocktail drop and dispensing the solution to each of the replicate experiments.
Step 3. Add 20 μL of Paraffin oil (PX0045‐3 EMD chemicals) to each well, after each row of 5 replicate experiments is completed.
Once all of the drops have been set up for a given plate, cover all of the wells in the plate with an additional 5‐6 mL of Paraffin oil. Incubate the plate at 23°C.