Resuspend DNA
This step uses RA1 to resuspend the precipitated DNA.
The RA1 volume used for DNA resuspension allows for samples to be hybridized at least in duplicate if needed. Samples are resuspended with 46 µl RA1 total, using 14 µl per sample inlet of the BeadChip.
About Reagents
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The RA1 volume used for DNA resuspension allows for samples to be hybridized at least in duplicate if needed. Samples are resuspended with 46µl RA1 total, using 26µl per sample inlet of the BeadChip. |
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Dispense only the volume necessary for each step. The Extend and Stain BeadChips procedure also requires RA1. |
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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. |
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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. Ventilation should be appropriate for handling of hazardous materials in reagents. 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, refer to the SDS at support.illumina.com/sds.html.
Preparation
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1.
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If the MSA4 plate was stored at -25°C to -15°C, thaw at room temperature, and then remove and discard the cap mat. |
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2.
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Prepare the following consumable: |
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RA1
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-25°C to ‑15°C
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Thaw at room temperature and invert to mix.
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3.
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Preheat the Illumina Hybridization Oven to 48°C. |
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4.
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Preheat the heat sealer for at least 20 minutes before use. |
Procedure
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1.
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Add 46 µl RA1 to each well of the MSA4 plate. |
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2.
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With the dull side facing down, apply a foil heat seal to the plate. Hold the heat sealer sealing block down firmly and evenly for 5 seconds. |
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3.
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Incubate in the preheated Illumina Hybridization Oven for 1 hour. |
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4.
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Vortex at 1800 rpm for 1 minute. |
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5.
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Pulse centrifuge at 280 × g. |
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6.
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Repeat steps 4 and 5 as needed to resuspend the pellets. |
SAFE STOPPING POINT
If you are stopping, store sealed MSA4 plate(s) at 2°C to 8°C for up to 24 hours. If more than 24 hours, store at -25°C to -15°C.
[Optional] If you are not stopping, you can set aside the MSA4 plate for up to 1 hour.
Store sealed RA1 at ‑25°C to ‑15°C. If using RA1 the next day, seal it, and store it overnight at 4°C.
