ZR-96 Soil Microbe DNA Kit™

Simple, efficient isolation of humic-free DNA from microbes in soil, sludge, sediment, sand, and water in minutes including tough-to-lyse bacteria, fungi, algae, and protozoa.
Ultra-high density BashingBeads™ are fracture resistant and chemically inert.

Product Size Catalog # Price Qty
ZR-96 Soil Microbe DNA Kit™ 2x96 Preps. D6002
$663.00

About ZR-96 Soil Microbe DNA Kit™

The ZR-96 Soil Microbe DNA Kit™ is designed for the simple and rapid isolation of humic-free, PCR-quality DNA from microbes in soil. This product can be used to isolate DNA from tough-to-lyse bacteria, fungi, protozoa, and algae that inhabit a variety of samples including clay, sandy, silty, peaty, chalky, and loamy soils. Soil microbes are rapidly and efficiently lysed by bead beating with our state of the art, ultra-high density BashingBeads™. Fast-Spin plate technology is then used to isolate the DNA, which is subsequently filtered to remove humic acids/polyphenols that can inhibit PCR. The procedure can be performed in minutes, and there is no need for organic denaturants or proteinases.


Format 96-well Plate
DNA Recovery Typically, up to 5 µg total DNA (per well) is eluted into ≥ 25 µl DNA Elution Buffer per sample.
Processing Time 50 min
Equipment Microcentrifuge, vortex, cell disruptor/pulverizer (optional).
DNA Size Limits Capable of recovering DNA fragments from 100 bp to ≥40 kb. Typical fragment sizes range from 25 kb "“ 35 kb. If present, parasitic and viral DNA will also be recovered.
Sample Sources DNA is isolated from bacteria, fungi, protozoa, and algae in up to 135mg of soil or from fungal/bacterial cells directly.
DNA Purity High-quality, humic-free DNA is eluted with DNA Elution Buffer and is especially well suited for PCR (A260/280 nm >1.8.

Metagenomic DNA isolated from 5 soil samples. M: 1 kb marker (NEB); 1-5: soil samples (sand, sandy clay loam, hydrophobic sandy loam, course sandy loam, fine gravel).

Step 1

Add up to 135 milligrams of soil sample to the tubes of a ZR BashingBead™ Lysis Rack.  Add 400 µl Lysis Solution to each tube.  Cap tubes tightly to prevent leakage.

Alternatively, add 10-20 mg (wet weight) fungal and/or bacterial cells that have been resuspended in up to 50 µl of water or isotonic buffer (e.g., PBS) to the tubes of a ZR BashingBead™ Lysis Rack.

Step 2

Secure in a 96-well block/plate bead beater (e.g., 2010 GenoGrinder®) and process samples.  Optimization of processing time/speed will be necessary for complete sample lysis.

Processing times may be as little as one minute when using high-speed cell disrupters (e.g., 2000 GenoGrinder®, page 5).  See manufacturer’s literature for operating information.

Step 3

Centrifuge the ZR BashingBead™ Lysis Rack at ≥ 3,000 x g (5,000 x g max.) for 5 minutes.

Step 4

Transfer up to 250 µl supernatant to each well of a 96-Well Block.

Step 5

Add 750 µl of Soil DNA Binding Buffer to the supernatant in the 96-Well Block from Step 4.  Cover completely with Cover Foil and mix thoroughly by vortexing for 2 minutes.  Centrifuge the 96-Well Block at ≥ 3,000 x g (5,000 x g max.) for 5 minutes.  

Step 6

Remove or pierce foil and transfer 500 µl of each of the supernatants from Step 5 to the wells of a Silicon-A™ Plate on a Collection Plate.  Centrifuge the assembly at ≥ 3,000 x g (5,000 x g max.) for 5 minutes.

Step 7

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

Step 8

Add 200 µl DNA Pre-Wash Buffer to the wells of the Silicon-A™ Plate on the emptied Collection Plate and centrifuge the assembly at ≥ 3,000 x g for 5 minutes.

Step 9

Add 500 µl Soil DNA Wash Buffer to the wells of the Silicon-A™ Plate on the Collection Plate and centrifuge the assembly at ≥ 3,000 x g for 5 minutes.

Step 10

Transfer the Silicon-A™ Plate onto a clean Elution Plate and add 100 µl (50 µl minimum) DNA Elution Buffer directly to the matrices in the plate. Centrifuge the assembly at ≥ 3,000 x g for 5 minutes.

If fungi or bacterial cultures were sampled, the DNA is now suitable for PCR as well as other downstream applications.

Step 11

Transfer the eluted DNA from Step 10 to a prepared Silicon-A™-HRC Plate mounted onto a clean Elution Plate.  Centrifuge the assembly at exactly 3,500 x g for 3 minutes. 

Eluted, ultra-pure DNA is now ready for use in your experiments, or the Elution Plate can be covered with Cover Foil for storage of the DNA.

 

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