Resuspend DNA (Manual)

This step uses IBX to resuspend the precipitated DNA.

• IBX
• Foil heat seals
• Use fresh IBX. Properly stored IBX that has not been used is considered fresh.
1. If the WGA plate was stored at -25°C to -15°C, thaw at room temperature, and then remove and discard the cap mat or foil adhesive seal.
2. Prepare the following consumable:

Reagent

Storage

Instructions

IBX

-25°C to ‑15°C

Thaw at room temperature and vortex or invert 10 times to mix. Centrifuge at 280 × g for 1 minute.
3. Preheat the Illumina Hybridization Oven to 58°C. Confirm temperature with NIST calibrated thermometer.
4. Preheat the heat sealer as follows.
• If you are using a programmable heat sealer, set to 165°C and 2.5 seconds.
• If you are using a manual heat sealer, switch it on for at least 20 minutes before use.
1. Add 46 µl IBX to each well of the WGA plate.
2. With the dull side facing down, apply a foil heat seal to the plate.
• If you are using a programmable heat sealer, hold down firmly and evenly for 2.5 seconds.
• If you are using a manual heat sealer, hold down firmly and evenly for 5 seconds.
3. Immediately remove the WGA plates from the heat sealer and firmly roll the rubber brayer over the plate until all well indentations are visible through the foil.
4. Incubate in the 58°C preheated Illumina Hybridization Oven for approximately 1.5 hours.
5. Remove plate from oven.
6. Vortex at 1800 rpm for 1 minute.
7. Pulse centrifuge at 280 × g to resuspend pellets.
8. [Optional] Unless you are stopping, you can set aside the WGA plate for up to 1 hour before proceeding.
9. Continue with Hybridize DNA to the BeadChip (IAPS with ILASS).

SAFE STOPPING POINT

If you are stopping, store the sealed WGA plates at -25°C to -15°C for up to 24 hours.

If stopping for longer than 24 hours, store at -80°C.