Precipitate DNA

This process begins with an isopropanol precipitation, and then it centrifuges to collect the fragmented DNA.

• PM1
• 100% 2-propanol
• Cap mats
1. If the MSA6 plate was stored at -25°C to -15°C, prepare it as follows:
a. Thaw at room temperature.
b. Pulse centrifuge at 280 x g for 1 minute.
c. Preheat the heat block to 37°C.
2. Prepare the following consumables:

Item

Storage

Instructions

PM1

2°C to 8°C

Thaw at room temperature and invert to mix. Centrifuge at 280 × g for 1 minute.
3. Remove the cap mat.
1. Add 100 μl PM1 to each well of the MSA6 plate.
2. Reseal with the cap mat using the original orientation.
3. Vortex the plate at 1600 rpm for 1 minute.
4. Incubate on the preheated heat block for 5 minutes.
5. Pulse centrifuge at 280 × g for 1 minute.
6. Set the centrifuge at 4°C in preparation for the next centrifuge step.
7. Remove and discard the cap mat.
8. Add 300 μl 100% 2-propanol to each sample well.
9. Carefully seal with a new, dry cap mat. Avoid shaking the plate until the mat is seated.
10. Invert the plate 10 times to mix.
11. Incubate in a refrigerator set at 4°C for 30 minutes.
12. Centrifuge at 3000 × g at 4°C for 20 minutes.
13. 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.
14. Make sure that a blue pellet is present in the bottom of each sample well.
15. Remove and discard the cap mat.
16. 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 smack the plate down on a dry area of the pad. Avoid the liquid drained onto the pad.
17. Keeping the plate inverted, firmly tap until all wells are free of liquid (~1 minute). Do not allow supernatant to pour in to other wells.
18. Place the uncovered, inverted plate on the tube rack for 1 hour at room temperature to air-dry the pellets.
19. Make sure that a blue pellet is still present in the bottom of each sample well.
20. 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.

Inverted MSA6 Plate

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

SAFE STOPPING POINT

If you are stopping, seal the plate, and store at -25°C to -15°C for up to 24 hours.