Promega Maxwell RSC Fecal Microbiome DNA Kit Instructions For Use

Maxwell RSC Fecal Microbiome DNA Kit
®
Instructions for Use of Product AS1700.

Preparing Fecal Microbiome Samples for DNA Purification

This Quick Protocol provides instructions for use of the Maxwell
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.
Additional protocol information is in Technical Manual #TM640, available online at: www.promega.com
® RSC Fecal Microbiome DNA Kit with the Maxwell
Quick Protocol
® Instruments to
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