TITLE: Quick-Start Protocol for DNeasy® PowerSoil® Pro Kit
AUTHORS: Carlos Goller

[DESCRIPTION]
This protocol will allow users to operate the DNeasy® PowerSoil® Pro Kit. 

[BEFORE_START]
Ensure that the PowerBead Pro Tubes rotate freely in the centrifuge without rubbing. 
If Solution CD3 has precipitated, heat at 60°C until the precipitate dissolves. 
Perform all centrifugation steps at room temperature (15–25°C).

[GUIDELINES]
Solution CD2 should be stored at 2–8°C upon arrival. All other reagents and kit components should be stored at room temperature (15–25°C). 

Further information
 DNeasy® PowerSoil® Pro Kit Handbook: www.qiagen.com/HB-2495
 Safety Data Sheets: www.qiagen.com/safety
 Technical assistance: support.qiagen.com

[STEPS]
SECTION: Using the DNeasy® PowerSoil® Pro Kit
1. Spin the PowerBead Pro Tube briefly to ensure that the beads have settled at the bottom. Add up to 250 mg  of soil and 800 µL  of Solution CD1. Vortex briefly to mix.

SECTION: Using the DNeasy® PowerSoil® Pro Kit
2. Secure the PowerBead Pro Tube horizontally on a Vortex Adapter for 1.5–2 ml tubes (cat. no. 13000-V1-24). Vortex at maximum speed for 10 min 

 

SECTION: Using the DNeasy® PowerSoil® Pro Kit
3. Centrifuge the PowerBead Pro Tube at 15000 x g  for 1 min 

SECTION: Using the DNeasy® PowerSoil® Pro Kit
4. Transfer the supernatant to a clean 2 ml Microcentrifuge Tube (provided)
 

SECTION: Using the DNeasy® PowerSoil® Pro Kit
5. Add 200 µL  of Solution CD2 and vortex for 5 s 

SECTION: Using the DNeasy® PowerSoil® Pro Kit
6. Centrifuge at 15000 x g  for 1 min at Room temperature . Avoiding the pellet, transfer up to 700 µL  of supernatant to a clean 2 ml Microcentrifuge Tube (provided)
 


SECTION: Using the DNeasy® PowerSoil® Pro Kit
7.  Add 600 µL of Solution CD3 and vortex for 5 s .

SECTION: Using the DNeasy® PowerSoil® Pro Kit
8. Load 650 µL  of the lysate onto an MB Spin Column and centrifuge at 15000 x g  for 1 min  

SECTION: Using the DNeasy® PowerSoil® Pro Kit
9. Discard the flow-through and repeat step 8 to ensure that all of the lysate has passed through the MB Spin Column.
SECTION: Using the DNeasy® PowerSoil® Pro Kit
10. 0. Carefully place the MB Spin Column into a clean 2 ml Collection Tube (provided).
 

SECTION: Using the DNeasy® PowerSoil® Pro Kit
11. Add 500 µL  of Solution EA to the MB Spin Column. Centrifuge at 15000 x g  for 1 min 

SECTION: Using the DNeasy® PowerSoil® Pro Kit
12. . Discard the flow-through and place the MB Spin Column back into the same 2 ml Collection Tube. 
SECTION: Using the DNeasy® PowerSoil® Pro Kit
13. Add 500 µL  of Solution C5 to the MB Spin Column. Centrifuge at 15000 x g  for 1 min 

SECTION: Using the DNeasy® PowerSoil® Pro Kit
14. Discard the flow-through and place the MB Spin Column into a new 2 ml Collection Tube (provided).
SECTION: Using the DNeasy® PowerSoil® Pro Kit
15. Centrifuge at up to 16000 x g  for 2 min . Carefully place the MB Spin Column into a new 1.5 ml Elution Tube (provided)

SECTION: Using the DNeasy® PowerSoil® Pro Kit
16. . Add 50 mL -100 mL  of Solution C6 to the center of the white filter membrane.

SECTION: Using the DNeasy® PowerSoil® Pro Kit
17. Centrifuge at 15000 x g  for 1 min . Discard the MB Spin Column. The DNA is now ready for downstream applications