Precipitate DNA (Manual)

•

100% 2-propanol (isopropanol)

•

PM1

•

Foil adhesive seal

•

Kimwipes

1.

If the WGA plate was stored at -25°C to -15°C, thaw at room temperature.

2.

Set the refrigerated centrifuge to 4°C.

3.

Prepare the following consumable:

1.

Vortex the plate at 1600 rpm for 1 minute, then centrifuge the plate at 280 × g for 1 minute.

2.

Remove and discard the cap mat.

3.

Add 100 μl PM1 to each well of the WGA plate.

4.

Add 300 μl 100% 2-propanol to each sample well.

5.

Carefully seal using a foil adhesive seal.

6.

Invert the plate 10 times to mix.

7.

Centrifuge at 3000 × g at 4°C for 20 minutes.

8.

Immediately remove the plate from the centrifuge.

•

When centrifuging is complete, proceed immediately to avoid dislodging the blue pellets.

•

If a delay occurs, repeat the 20-minute centrifuge.

9.

Make sure that a blue pellet is present in the bottom of each sample well.

10.

Remove and discard the foil adhesive seal.

11.

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.

12.

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.

13.

Keeping the plate inverted, firmly tap until all wells are free of liquid (~1 minute). Do not allow supernatant to pour into other wells.

14.

Place the uncovered, inverted plate on the tube rack for 40–45 minutes at room temperature to air-dry the pellets. Do not exceed 1-hour dry time.

15.

Make sure that a blue pellet is still present in the bottom of each sample well.