About Xpedition™ Plant/Seed DNA MiniPrep
Note: This product will be discontinued within the next 3 months (November 30th, 2016) or while supplies last. Please refer to ZR Plant/Seed DNA MiniPrep (D6020) 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™ Plant/Seed 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™ Plant/Seed DNA MiniPrep is designed for the simple, rapid isolation of inhibitor-free, PCR-quality DNA from a variety of plant sample sources including leaves, stems, buds, flowers, fruit, seeds, etc. 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 plant 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 polyphenol, lipid, polysaccharide-free DNA from the lysates. Eluted DNA is ideal for PCR for genotypic analysis including metagenomics, barcoding, phylogenetics, forensics, 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 50-100 µl (25 µl minimum) DNA Elution Buffer per sample.|
|Equipment||Microcentrifuge, vortex, cell disrupter/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||Up to 150 mg that include leaves, stems, buds, flowers, fruit, seeds, etc.|
|DNA Purity||High quality, inhibitor-free DNA is eluted with DNA Elution Buffer that is suitable for PCR amplification. A260/A280 < 1.8|
|DNA Yield||Typically 20-80 ng DNA/mg plant material.|
Add up to 150 mg of finely cut plant or seed sample to a ZR BashingBead™ Lysis Tube. Add 750 µl Xpedition™ Lysis/Stabilization Solution to the tube.
Note: The Xpedition™ Lysis/Stabilization Solution can be added to the ZR BashingBead™ Lysis Tube ahead of time, prior to sample addition
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 Plant/Seed DNA Binding Buffer to the filtrate in the Collection Tube from Step 4 and mix.
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 Plant/Seed 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 50-100 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.