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AQUASTOOL

AquaStoolTM is a multifunctional aqueous solution-based reagent for fecal DNA and RNA preservation and extraction, and fecal PCR inhibitor removal. It may be used to extract fecal DNA for non-invasive genotyping of transgenic animals. Unlike tail/toe/ear snipping, fecal sampling is not limited by animal age, physiopathological condition, and sampling frequency. Fecal DNA is particularly useful for verifying adult animals’ genotype and transgene stability. AquaStool may also be used to stabilize and extract fecal DNA/RNA from human stools for host and microbiome studies.


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Features
 

Multifunctionalities

AquaStool combines the functions of specimen stabilization, DNA extraction, and PCR inhibitor removal in a single solution.  

 

Non-invasive genotyping

 

AquaStool can be used to extract fecal DNA and enables noninvasive genotyping of transgenic animals and monitoring of wildlife animals. 

 

Recover DNA from the host and microbes in the stool

 

AquaStool extracts fecal DNA and RNA originated from the host, commensal bacteria, invasive viruses, fungi, or parasites, and incompletely digested foods, enabling the study of human and animal microbiome; diagnosis of bacterial, viral, fungal, and parasitic infections.

 

Not only for extraction but also for specimen stabilization

AquaStool may be used to collect fecal specimens in the field. It will stabilize the DNA and allow shipping the fecal specimens to the laboratory at ambient temperature.


Comparison

We offer AquaStool for fecal DNA extraction and AquaGenomic for tail DNA extraction. Here is a comparison of the two methods. 

   Comparison of tail DNA and fecal DNA extraction
 

Tail DNA

Fecal DNA

Reagent AquaGenomic * AquaStool

Specimen

Tail or ear snip

Fecal pellet

Invasive

Yes

No

Repeated sampling

Not allowed

Allowed

Age limitation

Under 21 days

Unlimited

Use lysate directly for PCR

Yes

No

Cost per extraction

$0.33 $1.00

GI microbial DNA

No

Yes


AquaGenomic (#2030) may also be used to extract fecal DNA from mice (not yet tested on other species). AquaGenomic is nontoxic and its lysate may be used directly for PCR.




Brief Protocol 

(Please refer to the Instruction Manual for fecal DNA/RNA extraction from human stool specimens.)

AquaStool mouse fecal DNA extraction protocol

1. Collect the fecal sample

Transfer a mouse to a cage floored with a clean paper towel. Scoop up a fecal pellet with a microfuge tube labeled with the animal ID (fecal pellets may be stored at RT for months after being air-dried by incubating in open tubes on a 37 °C heat bloc for 24 hrs).

2. Extract the DNA

Add 150 ul AquaStool (30 ml in the kit for 200 extractions) to each fecal pellet. Soak the pellet until it becomes soften and vortex vigorously to homogenize the fecal material (if needed, use a pipette tip to break up the fecal pellet). For microbial DNA extraction, vortex the fecal material in the presence of white sand or homogenize it with a bead beater.

3. Remove the Debris

Centrifuge the sample at 14,000 xg for 10 min to pellet the debris. 

4. Pellet the DNA

Transfer the supernatant (~90 ul) to a new 0.5-ml microfuge tube. Add 0.8 vol (~72 ul) of isopropanol and vortex to mix. Centrifuge at 14,000 xg for 5 min to pellet the DNA. Decant to discard the supernatant and rinse the DNA pellet with 70% ethanol twice. Suspend the DNA pellet in 100 ul of TE buffer or deionized water. Centrifuge again to pellet any insoluble material and transfer the DNA solution to a new tube (Note: It is critical to centrifuge the DNA solution again before adding it to a PCR reaction, as some insoluble material, which contain PCR inhibitors, may develop during storage.).


AquaGenomic mouse fecal DNA extraction protocol


1. Collect the fecal pellet

Transfer a mouse to a cage floored with a clean paper towel. Scoop up a fecal pellet with a microfuge tube labeled with the animal ID (fecal pellets may be stored at RT for months after being air-dried by incubating in open tubes on a 37 °C heat bloc for 24 hrs).

2. Extract the DNA

Add 200 ul of AquaGenomic solution to each fecal pellet. Incubate at 85 °C for 20 min. (For tail DNA extraction, place the tissue into a microfuge tube preloaded with 100 ul of AquaGenomic solution containing 10 ug of Proteinase K. Incubate at 60 °C for 2 hrs and then at 95 °C for 10 minutes to inactivate the Proteinase K. Homogenize the tissue by vortexing or pipetting.)

3. Pellet the Debris

Centrifuge at 12,000 xg for 4 min to pellet the debris. Transfer the supernatant (~100 ul) to a new 0.5-ml microfuge tube. (Note: To use the lysate for PCR, dilute an aliquot of the lysate with 9 vol of deionized water and use 0.5 ul of the diluted lysate in a 25-ul PCR reaction.)

4. Pellet the DNA

Add 0.8 vol (~80 ul) of isopropanol and mix by vortexing for 30 sec. Centrifuge at 12,000 xg for 2 min to pellet the DNA. Flip the tube to discard the supernatant. Fill the microfuge tube with 70% ethanol by shooting the ethanol solution at the cap of the tube from a squirt bottle, and then flip to discard the ethanol. Repeat the 70% ethanol rinse 2 times. Air-dry the DNA pellet. Add 50 ul of TE buffer or deionized water, vortex vigorously to suspend the DNA. Centrifuge again for 2 min to pellet any insoluble and transfer the clear DNA solution to a new tube.

 

 

Ordering Information


Product Name:    AquaStool Kit

Product Number:   7001, 7030

Application:   DNA/RNA extraction from fecal specimens. For in vitro research use only.

Size:    The kit is sufficient for the preparation of

      • 7001: 6 extractions (use 150 ul AquaStool for a mouse fecal pellet)
      • 7030: 200 extractions 

Kit Contents:   The AquaStool Kit includes the following items

      • 7001: 1 ml AquaStool Solution, Instruction Manual
      • 7030: 30 ml AquaStool Solution, Instruction Manual

Price:  

      • 7001: $10 each
      • 7030: $199 each

Ordering:    To order, please click the "BUY" button below.

  

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MultiTarget Pharmaceuticals, LLC - All rights reserved