Promega Maxwell AS1831 User manual

Quick Protocol
Instructions for Use of Product AS1831.
Maxwell®RSC Enviro Total Nucleic Acid Kit
Wastewater sample (40ml).
1. Add Binding Buffer 1 to each tube.
2. Add Binding Buffer 2 to each tube.
3. Add isopropanol to each tube. Mix.
Add Protease Solution, centrifuge,
then divide supernatant
equally into two tubes.
Transfer liquid to Reservoir Extension
Funnel on PureYield™ Binding Column.
Apply vacuum.
Place microcentrifuge tube in
Eluator™ Device, add Nuclease-Free
Water and apply vacuum to elute
nucleic acid.
1. Add Column Wash 1, apply vacuum.
2. Add Column Wash 2, apply vacuum.
17691MA
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 × gfor 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)
PAGE 1 •PART #FB237
Figure 1. Schematic showing direct capture of nucleic acid
from wastewater using a PureYield™ Binding Column.

1. .
Quick Protocol
Instructions for Use of Product AS1831.
Maxwell®RSC Enviro Total Nucleic Acid Kit
PROMEGA CORPORATION •2800 WOODS HOLLOW ROAD •MADISON, WI 53711-5399 USA •TELEPHONE [phone]
www.promega.com •© 2021 PROMEGA CORPORATION •ALL RIGHTS RESERVED •PRICES AND SPECIFICATIONS SUBJECT TO CHANGE WITHOUT PRIOR NOTICE •PRINTED IN USA 07/21 •PART #FB237
11339TA
1
23
45
67
8
1. Binding Buffer
2. Magnetic Particles
1. Preprocessed sample
8. CSC Plunger
3. Wash Solution
4. Wash Solution
5. Wash Solution
6. Wash Solution
7. Empty
8. Empty
Well Content User Adds:
Additional protocol information in Technical Manual #TM663, available online at: www.promega.com
Total Nucleic Acid Extraction and Clean-Up on Maxwell®
Instrument
Cartridge Preparation
1. Place the cartridge to be used in the Deck Tray(s) with well #1 (the largest well)
facing away from the elution tube.
2. Press down on the cartridge to snap it into position. Carefully peel back the seal
so that all plastic comes off the top of the cartridge. Ensure that all sealing tape
and any residual adhesive are removed before placing cartridges in the
instrument.
3. Place a plunger in well #8 of each cartridge. Well #8 is the well closest to the
elution tube.
4. Place an empty elution tube in the elution tube position for each cartridge. Add 80μl of Nuclease-Free
Water to the bottom of each elution tube.
5. Add 150μl of Binding Buffer 1 and 50μl of Binding Buffer 2 to the 0.5ml of liquid eluted in Step 18.
6. Load the entire volume of mixture to well #1 (the largest well).
Maxwell®Instrument Run
Follow the instrument run instructions in the Maxwell
®
RSC Enviro Total Nucleic Acid Kit Technical Manual, #TM663.
Figure 2. Maxwell®RSC Cartridge. Sample concentrate is
added to well #1, plunger added to well #8.