Precipitate DNA

This step uses 100% 2-propanol and PM1 to precipitate the DNA.

• 100% 2-propanol
• PM1
• Foil adhesive seal
• Kimwipes
1. If the WGA plate was stored at -25°C to -15°C, thaw to room temperature, vortex at 1600 rpm for 1 minute, and then centrifuge at 280 × g for 1 minute.
2. Remove and discard the cap.
3. Prepare the following consumables:

Reagent

Storage

Instructions

PM1

2°C to 8°C

Thaw at room temperature. Vortex or invert 10 times to mix. Centrifuge reagent tubes at 280 x g for 1 minute, or flick bottle to remove reagent from lid.
4. Prepare the required amount of PM1 and isopropanol reagent according to instructions on the interface.

Number of samples

PM1

Isopropanol (ml)

48

2 tubes

35

96

2 tubes

50

192

4 tubes

90

288

6 tubes

120

384

1 60-ml bottle

160

480

1 60-ml bottle

200

576

2 60-ml bottle

240
5. Set refrigerated centrifuge to 4°C.
1. Open ILASS on the Infinium Automated Pipetting System.
2. If running with PGx, select Infinium Genotyping Assay with PGx, then Precipitate DNA.

If running without PGx, select Infinium Genotyping Assay, then Precipitate DNA.

3. Select the Product Type drop-down, and then select your product type.
4. In the Container Input Quantity box, enter the number of 96-well plates to be processed. Manually enter the value or set the value using +/-.

ILASS displays the number of required materials based on number of plates.

5. Select Continue.
6. Setup the Infinium Automated Pipetting System worktable as shown on the interface, according to number of plates used in the run.
7. Before loading the worktable, mix PM1 by inverting 10 times and centrifuging at 280 x g for 1 minute for reagent tubes, or flick 60 ml bottle to remove reagent from lid.
8. Pour PM1 reagent into the PM1 half-trough.
9. Pour the required amount of isopropanol into 2-propanol full-trough.
10. Select Auto-Scan Item(s).
11. Manually enter information for trough reagents by following the user interface.
12. After successful scanning, the user interface displays a timer.
13. Select Execute Run.
14. Wait for the Infinium Automated Pipetting System to finish the run.
15. Seal the WGA plates with foil adhesive seal.
16. Remove troughs from the worktable. Discard residual reagent volume. Clean troughs with DI water.
17. Invert the plate 10 times to mix.
18. Centrifuge at 3000 × g at 4°C for 20 minutes.
• When centrifuging is complete, proceed immediately to the next step to avoid dislodging the blue pellet.
• If a delay occurs, repeat the 20 minute centrifuge.
19. Remove WGA plate from centrifuge.
20. Make sure that a blue pellet is present in the bottom of each sample well.
21. Remove and discard the foil adhesive seal.
22. Hold the plate over an absorbent pad and do as follows.
a. Quickly invert to decant the supernatant.
b. Drain liquid onto the absorbent pad, and then firmly tap the plate down on a dry area of the pad.
c. Keeping the plate inverted, use a Kimwipe to remove any residual alcohol on the surface of the plate or draining from the wells of the plate.
d. Keeping the plate inverted, firmly tap until all wells are free of liquid (~1 minute). Do not allow supernatant to pour into other wells.
e. Place the uncovered, inverted plate on the tube rack for 40 minutes at room temperature to air-dry the pellets. Do not exceed 1 hour dry time.
f. Make sure that a blue pellet is still present in the bottom of each sample well.

Inverted WGA Plate

Do not overdry the pellets. Pellets that are overdried are difficult to resuspend and can lead to poor data quality.