About Xpedition™ Soil/Fecal DNA MiniPrep
Note: This product will be discontinued within the next 3 months (November 30th, 2016) or while supplies last. Please refer to ZR Fecal DNA MiniPrep (D6010) for the replacement product; the Xpedition Stabilization/Lysis Solution (D6202-1-40) can be purchased separately.Degradation and contamination of biological samples have been obstacles to scientific study, and may be particularly problematic in highly sensitive molecular-analysis techniques (e.g., PCR of low copy DNA). Use of cryogenic freezing methods for environmental/forensic sample preservation may often be too impractical to be employed. The Xpedition™ Soil/Fecal DNA MiniPrep, when used with a portable bead beating device [i.e., Xpedition™ Sample Processor (XSP), pg. 6], allows the researcher/investigator to "Take the Lab to the Field" and to the site of sample collection. The Xpedition™ Soil/Fecal DNA MiniPrep can be used for the isolation of inhibitor-free DNA from tough-to-lyse bacteria, fungi, protozoa, algae, etc., that inhabit a range of samples including feces as well as clay, sandy, salty, silty, peaty, chalky, and loamy soils. Samples from remote locations can be processed at the site of collection using the provided ZR BashingBead™ Lysis Tubes and a XSP (available separately, Cat. No. S6020, pg. 6). Sample processing occurs in a unique lysis/stabilization solution that effectively preserves DNA in soil/fecal lysates for subsequent storage and transport. Later, when convenient with the researcher, the kit's Fast-Spin column technologies are used for the isolation of humic acid/polyphenol-free DNA from the lysates. This product can also be used for isolation of DNA from cultured bacteria, fungi, and yeast. Eluted DNA is ideal for PCR for various genotypic analysis including metagenomics, forensics, barcoding, phylogenetics, etc. An overview of the procedure is shown below.
|Format||Bead Beating, Spin Column|
|DNA Recovery||Typically, up to 25 µg total DNA is eluted into 100 µl (25 µl minimum) DNA Elution Buffer per sample.|
|Equipment||Microcentrifuge, Vortex, Portable Disruptor/Bead Beater (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||Fungi, protozoa, algae, etc. in up to 0.25 g of soil, feces, dirt, sediment, and sludge|
|DNA Purity||High quality, polyphenolic-free DNA is eluted with DNA Elution Buffer making it perfect for PCR. A260/A280 <1.8|
Add up to 0.25 grams of soil or fecal material sample to a ZR BashingBead™ Lysis Tube. Add 750 µl Xpedition™ Lysis/Stabilization Solution to the tube.
Alternatively, add 50-100 mg (wet weight) fungal and/or bacterial cells that have been resuspended in up to 200 µl of water or isotonic buffer (e.g., PBS) to a ZR BashingBead™ Lysis Tube.
Secure in a bead beater fitted with a 2 ml tube holder assembly (e.g., Xpedition™ Sample Processor for processing at the site of sample collection) and process for a minimum of 30 seconds. Store/transport samples. Proceed to Step 3 when convenient.
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 x g (~7,000 rpm) for 1 minute.
Add 1,200 µl of Soil/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 Soil/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.
If fungi or bacterial cultures were sampled, the DNA is now suitable for PCR as well as other downstream applications.
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.