DNA extraction from fecal samples

Place 50-100mg of fecal sample into tube.

Add 1mL of PBS per 100mg of feces.

Mix thoroughly by vortexing.

Allow the sample to stand for 0h 2m 0s to sediment large debris.

Transfer 300µL of the suspension to 1.5 mL tube.

Centrifuge at 10000x g.

Discard the supernatant.

Resuspend the pellet (~30mg of feces) in 300µL of PBS.

Incubate at 70°C for 0h 10m 0s on the block heater.

Cool to Room temperature.

Transfer 300µL of the suspension to EZ-beads tube.

Lyse cells either by using disruption device (12.1) or vortex mixer (12.2).

Place the EZ-beads tube in Micro Smash instrument and disrupt cells by shaking at 2500rpm.

Equipment

Value	Label
Micro Smash Beads Cell Disrupter	NAME
TOMY Digital Biology	BRAND
MS-100	SKU

Place the EZ-beads tube on MN Bead Tube Holder attached to Vortex-Genie mixer and vortex for 0h 5m 0s at maximum speed.

Equipment

Value	Label
MN Bead Tube Holder	NAME
Rubber-foam adapter for processing bead tubes with Vortex-Genie instrument	TYPE
MACHEREY-NAGEL	BRAND
740469	SKU
http://www.mn-net.com/	LINK

Briefly spin the tube to collect contents.

Add 300µL of Lysis Buffer and 30µL of Proteinase K Solution to the sample in EZ-beads tube.

Mix by inverting the tube.

Briefly spin the tube.

Incubate at 56°C for 0h 20m 0s on the block heater.

Briefly spin the tube.

Transfer the supernatant (~500µL ) to 1.5 mL tube.

Centrifuge at 18000x g.

Transfer the cleared lysate to Maxwell RSC Cartridge.

Add 50µL of Elution Buffer to elution tube.

Start the extraction run following the manufacturer's instructions.

Equipment

Value	Label
Maxwell RSC instrument	NAME
Automated nucleic acid purification platform	TYPE
Promega	BRAND
AS4500	SKU
http://www.promega.com	LINK