Promega Maxwell AS1831 Instructions For Use

Maxwell RSC Enviro Total Nucleic Acid Kit
®
Instructions for Use of Product AS1831.
Standard Protocol for Capture, Concentration and
Clean-Up

Direct Capture and Concentration

1. Add 40ml of pasteurized wastewater into a 50ml conical tube.
2. Add 0.5ml of Protease Solution. Mix by inversion and incubate for 30 minutes
at ambient temperature.
3. Clarify sample by centrifuging at 3,000 × g for 10 minutes.
4. Carefully decant 20ml of the supernatant into each of two new 50ml conical
tubes.
5. To each tube containing 20ml of the clarified supernatant, add 6ml of Binding
Buffer 1 (BBD) followed by 0.5ml of Binding Buffer 2 (BBE).
6. Mix well by inversion.
7. Add 24ml of isopropanol to each tube. Mix well by inversion.
8. Attach a Reservoir Extension Funnel to the PureYield™ Binding Column, then
connect the column to the vacuum manifold.
9. Pour the mixture from each tube from Step 7 into the Reservoir Extension
Funnel on the PureYield™ Binding Column.
10. Turn on the pump and apply vacuum to capture total nucleic acid on the
column.
11. Add 5ml of Column Wash 1 (CWE) and apply a vacuum to pull the liquid
through the PureYield™ Binding Column.
12. Add 20ml of Column Wash 2 (RWA) and apply a vacuum to pull the liquid
through the PureYield™ Binding Column.
13. Continue the vacuum for an additional 30 seconds after all liquid has passed
through the membrane.
14. Release the vacuum and remove the column from the vacuum manifold.
15. Assemble the elution device by placing a 1.5ml microcentrifuge tube into the
base of the Eluator™ Vacuum Elution Device.
16. Place the Eluator™ Device assembly onto a vacuum manifold.
17. Add 250μl of preheated (60°C) Nuclease-Free Water to the PureYield™ Binding
Column. Apply maximum vacuum until all liquid has passed through the column.
18. Repeat the elution by adding another 250μl of preheated Nuclease-Free Water to the
PureYield™ Binding Column.
(continued)
Quick Protocol
Wastewater sample (40ml).
Add Protease Solution, centrifuge,
then divide supernatant
equally into two tubes.
1. Add Binding Buffer 1 to each tube.
2. Add Binding Buffer 2 to each tube.
3. Add isopropanol to each tube. Mix.
Transfer liquid to Reservoir Extension
Funnel on PureYield™ Binding Column.
Apply vacuum.
1. Add Column Wash 1, apply vacuum.
2. Add Column Wash 2, apply vacuum.
Place microcentrifuge tube in
Eluator™ Device, add Nuclease-Free
Water and apply vacuum to elute
nucleic acid.
Figure 1. Schematic showing direct capture of nucleic acid
from wastewater using a PureYield™ Binding Column.
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