Resuspend DNA

This step uses RA1 to resuspend the precipitated DNA.

Consumables

• RA1
• Foil heat seals

About Reagents

• Dispense only the volume necessary for each step. The Extend and Stain BeadChips procedure also requires RA1.
• Use fresh RA1 for each step that requires it. Properly stored RA1 that has not been dispensed for this resuspension step or the extend and stain step is considered fresh.
• RA1 might form visible precipitate or crystals. Before each use, hold in front of a light and inspect. Invert several times to redissolve the solution as needed.

This set of reagents contains potentially hazardous chemicals. Personal injury can occur through inhalation, ingestion, skin contact, and eye contact. Wear protective equipment, including eye protection, gloves, and laboratory coat appropriate for risk of exposure. Handle used reagents as chemical waste and discard in accordance with applicable regional, national, and local laws and regulations. For additional environmental, health, and safety information, see the SDS at support.illumina.com/sds.html.

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:

Item

Storage

Instructions

RA1

-25°C to ‑15°C

Thaw at room temperature and invert to mix.
3. Preheat the Illumina Hybridization Oven to 48°C.
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, set to 165°C for at least 20 minutes before use.

1. Add 46 µl RA1 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 plate from the heat sealer and firmly roll the rubber plate sealer over the plate until all 96 well indentations are visible through the foil. Repeat application of heat sealer until all 96 wells are defined.
4. Incubate in the preheated Illumina Hybridization Oven for 1 hour.
5. Vortex at 1800 rpm for 1 minute.
6. Pulse centrifuge at 280 × g.
7. Repeat steps 5 and 6 as needed to resuspend the pellets.
8. [Optional] Unless you are stopping, you can set aside the WGA plate for up to 1 hour before proceeding.