Promega Maxwell RSC Fecal Microbiome DNA Kit User manual

Quick Protocol
Instructions for Use of Product AS1700.
Maxwell®RSC Fecal Microbiome DNA Kit
Additional protocol information is in Technical Manual #TM640, available online at: www.promega.com PAGE 1 •PART# FB234
Preparing Fecal Microbiome Samples for DNA Purification
This Quick Protocol provides instructions for use of the Maxwell®RSC Fecal Microbiome DNA Kit with the Maxwell®Instruments to
purify DNA human or mouse fecal samples. For detailed instructions, including information on instrument setup and troubleshooting,
please refer to the Maxwell®RSC Fecal Microbiome DNA Kit Technical Manual #TM640.
Note: To use the Maxwell®RSC Fecal Microbiome DNA Kit, you must have the “Fecal Microbiome DNA” method loaded on the
Maxwell®Instrument.
Materials to Be Supplied By User
• microcentrifuge tubes, 2.0ml
• sterile, aerosol-resistant pipette tips
• heat block
• microcentrifuge
• vortex
Preprocessing Fecal Samples
1. Place 100–300mg of fecal sample into a 2ml screw-cap microcentrifuge tube.
2. Add 1ml of Lysis Buffer and 40μl of Proteinase K to the microcentrifuge tube and vortex for 30 seconds.
3. Place the tube into a heat block at 95°C for 5 minutes.
4. Remove samples from heat block and allow to cool for 2 minutes on bench top.
5. Vortex thoroughly for 1 minute.
6. Incubate samples at 56°C for 5 minutes.
7. During the incubation (Step 6) prepare cartridges. See Preparing the Cartridge.
8. After incubation place lysate tubes into a microcentrifuge and spin at room temperature for 5 minutes at maximum speed
(>10,000 × g) to pellet the solids.
9. Transfer only 300μl of supernatant into well # 1 of the reagent cartridge (Figure 1). Avoid pipetting any solid material from the
bottom of the tube or on the surface of the liquid. Transferring these materials can inhibit downstream assays. If necessary, transfer
the supernatant to a new tube and centrifuge again to avoid solids.
Note: Some lysate will remain in the tube after transferring the 300μl aliquot to the cartridge.

Quick Protocol
Instructions for Use of Product AS1700.
Maxwell®RSC Fecal Microbiome DNA Kit
PROMEGA CORPORATION •2800 WOODS HOLLOW ROAD •MADISON, WI 53711-5399 USA •TELEPHONE [phone]
www.promega.com •© 2020 PROMEGA CORPORATION •ALL RIGHTS RESERVED •PRICES AND SPECIFICATIONS SUBJECT TO CHANGE WITHOUT PRIOR NOTICE •PRINTED IN USA •10/20 •PART# FB234
Additional protocol information is in Technical Manual #TM640, available online at: www.promega.com
Maxwell is a registered trademark of Promega Corporation.
Automated DNA Purification
Preparing the Cartridge
1. Place the cartridges to be used in the deck tray(s) with
well #1 (the largest well) facing away from the elution
tube (Figure 1).
2. Press down on the cartridge to snap it into position.
Carefully peel back the seal so the entire seal is
removed from the top of the cartridge. Ensure that all
sealing tape and any residual adhesive are removed
before placing cartridges in the instrument.
Caution: Handle cartridges with care. Seal edges may
be sharp.
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 into the Elution Tube
position for each cartridge in the deck tray(s). Ensure
that the caps are open and facing away from the
cartridge positions
5. Add 100μl of Elution Buffer to the bottom of each
Elution Tube.
Notes:
a. If Elution Buffer is on the side of the tube, elution may be suboptimal.
b. Use only the 0.5ml Elution Tubes provided in the kit; other tubes may not work with the Maxwell®RSC Instrument.
6. Add 300μl of Binding Buffer to well #1 (the largest well) of each cartridge.
7. Add 20μl of RNase A to well #3 of each cartridge.
8. Return to Step 8 of Preprocessing Fecal Samples.
Starting a Run on Maxwell®Instruments
Follow the instrument set up and run instructions in the Maxwell®RSC Fecal Microbiome DNA Kit Technical Manual #TM640.
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Figure 1. Set up and configuration of the deck tray.