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Xpedition™ Fungal/Bacterial DNA MiniPrep

"Take the Lab to the Field" and to the site of sample collection with state of the art Xpedition™ Sample Prep and DNA preservation technologies.
For efficient BashingBead™ lysis and isolation of inhibitor-free, PCR-quality DNA from tough-to-lyse fungi (e.g, yeast) and bacteria.
Unique lysis/stabilization solution keeps DNA intact in lysates for storage and transport following remote sample processing with the Xpedition™ Sample Processor (XSP).

Product Size Catalog # Price Qty
Xpedition™ Fungal/Bacterial DNA MiniPrep 50 Preps. D6206

About Xpedition™ Fungal/Bacterial DNA MiniPrep

Note: This product will be discontinued within the next 3 months (November 30th, 2016) or while supplies last. Please refer to ZR Fungal/Bacterial DNA MiniPrep (D6005) 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™ Fungal/Bacterial 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™ Fungal/Bacterial DNA MiniPrep is designed for the simple, rapid isolation of DNA from tough-to-lyse fungi, including A. fumigates, C. albicans, N. crassa, S. cerevisiae, S. pombe, as well as from mycelium and Gram (+) and (-) bacteria. 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 fungal/bacterial 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 inhibitor-free DNA from the lysates. Eluted DNA is ideal for PCR for various genotypic analyses including metagenomics, barcoding, phylogenetics, forensics, etc. An overview of the procedure is shown below.

Format Bead Beating, Spin Column Purification
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 50-100 mg (wet weight) fungi or bacteria or up to 200 mg tissue. This equates to approximately 109 bacterial cells, 108 yeast cells and 107 mammalian cells. Spores, pollen, nematodes, as well as other microorganisms can also be sampled.
DNA Purity High quality, polyphenolic-free DNA is eluted with DNA Elution Buffer making it perfect for PCR. A260/A280 <1.8

Step 1

Add 50-100 mg (wet weight) fungal or bacterial cells that have been resuspended in up to 200 µl of water or isotonic buffer (e.g., PBS) or up to 50 mg of tissue 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.

Step 2

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.

Step 3

Centrifuge the ZR BashingBead™ Lysis Tube in a microcentrifuge at 10,000 x g for 1 minute.

Step 4

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.

Step 5

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.

Step 6

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.  

Step 7

Discard the flow through from the Collection Tube and repeat Step 6.

Step 8

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.  

Step 9

Add 500 µl Fungal/Bacterial DNA Wash Buffer to the Zymo-Spin™ IIC Column and centrifuge at 10,000 x g for 1 minute.

Step 10

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.

Ultra-pure DNA is now ready for use in your experiments.

For specific notes and additional information, please see the product protocol PDF.
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