About ZR Fecal DNA MiniPrep™
The ZR Fecal DNA MiniPrep™ is designed for the simple and rapid isolation of inhibitor-free, PCR-quality host cell and microbial DNA from a variety of sample sources including humans, birds, rats, mice, cattle, etc. The procedure is easy and can be completed in minutes: fecal samples are rapidly and efficiently lysed by bead beating with our state of the art, ultra-high density BashingBeads™. Fast-Spin column technology is then used to isolate the DNA which is subsequently filtered to remove humic acids/polyphenols that can inhibit PCR. Eluted DNA is ideal for downstream molecular-based applications including PCR, arrays, genotyping, methylation detection, etc.
|DNA Recovery||Up to 25 μg total DNA is eluted into ≥ 25 μl DNA Elution Buffer per sample.|
|Processing Time||15 min|
|Equipment||Microcentrifuge, vortex, cell disruptor/pulverizer (optional)|
|DNA Size Limits||Capable of recovering genomic DNA up to and above 40 kb. In most instances, mitochondrial DNA and viral DNA (if present) will also be recovered.|
|Sample Sources||Bacterial, protist, as well as host DNA can be effectively isolated from a ≤150 mg sample of mammalian feces.|
|DNA Purity||High quality, inhibitor-free DNA is eluted with DNA Elution Buffer suitable for the amplification of bacterial, protist, and/or mammalian templates. A260/A280 >1.8|
Comparison of DNA yields from rat feces using the ZR Fecal DNA MiniPrep™ and kits from suppliers A and B. Equivalent amounts of feces were processed using each kit and then equal volumes of eluted DNA were analyzed in a 0.8% (w/v) agarose/ethidium bromide gel. Samples were processed in triplicate.
Add up to 150 mg of fecal sample to a ZR BashingBead™ Lysis Tube. Add 750 µl Lysis Solution to the tube.
Secure in a bead beater fitted with a 2 ml tube holder assembly (e.g., Disruptor Genie™) and process at maximum speed for 5 minutes.
Processing times may be as little as 40 seconds when using high-speed cell disrupters (e.g., the portable Xpedition™ Sample Processor, page 6, FastPrepâ-24, or similar). See manufacturer’s literature for operating information.
Centrifuge the ZR BashingBead™ Lysis Tube in a microcentrifuge at ≥10,000 x g for 1 minute.
Transfer up to 400 µl supernatant to a Zymo-Spin™ IV Spin Filter (orange top) in a Collection Tube and centrifuge at 7,000 rpm (~7,000 x g) for 1 minute.
Add 1,200 µl of Fecal DNA Binding Buffer to the filtrate in the Collection Tube from Step 4.
Transfer 800 µl of the mixture from Step 5 to a Zymo-Spin™ IIC Column in a Collection Tube and centrifuge at 10,000 x g for 1 minute.
Discard the flow through from the Collection Tube and repeat Step 6.
Add 200 µl DNA Pre-Wash Buffer to the Zymo-Spin™ IIC Column in a new Collection Tube and centrifuge at 10,000 x g for 1 minute.
Add 500 µl Fecal DNA Wash Buffer to the Zymo-Spin™ IIC Column and centrifuge at 10,000 x g for 1 minute.
Transfer the Zymo-Spin™ IIC Column to a clean 1.5 ml microcentrifuge tube and add 100 µl (25 µl minimum) DNA Elution Buffer directly to the column matrix. Centrifuge at 10,000 x g for 30 seconds to elute the DNA.
Transfer the eluted DNA from Step 10 to a prepared Zymo-Spin™ IV-HRC Spin Filter (green top) (see above) in a clean 1.5 ml microcentrifuge tube and centrifuge at exactly 8,000 x g for 1 minute. The filtered DNA is now suitable for PCR and other downstream applications.