FlyChip protocol for adding spotting buffer to 384-well printing plates
Overview
Oligonucleotide libraries are dispatched to us in 384-well microtitre plates. Spotting buffer is added to these plates using a Beckman Coulter Biomek NXP Liquid Handling Robot (LHR).
Equipment and reagents
- Beckman Coulter Biomek NXP Liquid Handling Robot (LHR)
- AP96 non-sterile P20 tips (Beckman Coulter; Cat. No. 717254)
- 70% Ethanol
- Spotting buffer
- Hettich Rotina 35 microtitre plate centrifuge
- Adhesive PCR Film (Abgene; Cat. No. AB-0558)
- Horizontal laminar flow work station (Jencons; Cat. No. 566-031)
Procedure
- Remove the microtitre plates from the -80 °C freezer and leave to defrost
- Once defrosted, centrifuge all plates at 2000 rpm for 2 minutes
- Clean the exterior and interior of the LHR using the Dyson vacuum cleaner and wipe with 70% Ethanol
- Open the program "Hydrate_LIBRARY_YesWash" and home all instrument drives
- Fill the in-flow wash tank with distilled water and prime the wash station, e.g., for 3-5 min.
- Load the instrument, as directed by the "Hydrate_LIBRARY_YesWash" program, i.e., fresh box of AP96 P20 tips, reservoir filled with spotting buffer and the plate to be hydrated
- Adhesive film should be removed just before the plate is loaded into the LHR
- All plates should be loaded in the LHR with well A1 in the top-left corner
- Start the program and watch to make certain the LHR is working correctly
- Repeat steps 6 to 7 until all plates have been rehydrated: as each plate is finished, remove from LHR and affix an adhesive PCR film
- Centrifuge all plates at 2000 rpm for 2 minutes and then incubate the plates at 37 °C for 2 hours to dissolved the probe DNA
- Clean the LHR to make certain that it has been left ready for others to use
- Centrifuge all plates at 2000 rpm for 2 minutes and store the plates at -80 °C
R. Auburn (24-02-2009).
