Resuspend DNA

This step uses IHX to resuspend the precipitated DNA.

• IHX
• Foil heat seals
• Use fresh IHX. Properly stored IHX that has not been used is considered fresh.
• Make sure that IHX is thawed at 37°C for at least 30 minutes prior to use. The thawing instructions are also included in Preparation.
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

IHX

-25°C to ‑15°C

Thaw at 37°C for at least 30 minutes prior to use. Vortex or invert 10 times to mix. Centrifuge at 280 x 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, set to 165°C for at least 20 minutes before use.
1. Add 46 µl IHXto 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. Repeat application of heat sealer until all 96 wells are defined.
4. Incubate in the preheated Illumina Hybridization Oven for approximately 1.5 hours.

During the incubation, in 30 minute intervals, perform the following:

a. Remove plate(s) from oven.
b. Vortex at 1800 rpm for 1 minute.
c. Pulse centrifuge at 280 x g to resuspend pellets.
d. Return to oven.
e. Repeat steps a - d every 30 minutes as needed to fully resuspend the pellets.