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AQUASTOOL

Stool is an accessible and underutilized noninvasive source of biospecimen. Fecal DNA/RNA originated from the host, intestinal bacteria, viruses, fungi, or parasites, and incompletely digested foods are molecular fingerprints of the host and its health. AquaStoolTM is a multifunctional aqueous solution-based reagent for fecal sample stabilization, DNA and RNA extraction and PCR inhibitor removal. It may be used to extract fecal DNA for non-invasive genotyping of transgenic animals. Fecal sampling is not limited by animal age, physiopathological condition, or sampling frequency. AquaStool may also be used to preserve and extract DNA/RNA from human stools for host and gut microbiota research.



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Features
 
 
Multi-functionalities
AquaStool combines the functions of specimen stabilization, DNA/RNA 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 host and microbial DNA and RNA in feces
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.

Powerful lysis
The powerful combination of AquaStool, sand and sonication lysis can produce 100-200 ug of DNA from 50 mg of feces. The sheared DNA is ready for next generation sequencing and could reveal a different but "true" microbiota profile of your samples.


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/RNA and allow the shipping of the fecal samples to the laboratory at ambient temperature, therefore, streamline fecal sample collection, transport and DNA/RNA extraction with a single reagent.


fDNA-RNA
      1   2   3   4

Figure 1. AquaStool purified fecal DNA and RNA.  Aliquots (5 μl) of the extracted DNA and RNA were electrophoresized in a native 0.8% agarose gel.

  1. DNA ladder.
  2. DNA extracted by vortexing.
  3. DNA/RNA extracted by sonication (The sheared DNA is ready for next generation sequencing. RNA contamination will not affect sequencing).
  4. DNase I treated RNA.


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 animal feces, where fecal PCR inhibitors are removed with the AquaPrecipi solution (#3015). If you are interested in mitochondrial DNA typing, we recommend the use of AquaGenomic for fecal DNA extraction.




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 mitochondrial DNA extraction, add Proteinase K to the AquaGenomic solution to 100 ug/ml, incubate the sample at 60 C for 2 hrs and then at 95 C for 10 minutes to inactivate the Proteinase K). Homogenize the heat-treated sample by vortexing or pipetting or bead-beating with a multichannel bead-beater.

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. 

4. Pellet the DNA

Add 0.5 vol (~50 ul) of AquaPrecipi (#3015, order separately) and 0.5 vol (~50 ul) of 95-100% of ethanol. Vortex for 60 sec and centrifuge at 12,000 xg for 5 min to pellet the DNA. Decant to discard the supernatant. Fill the tube with 70% ethanol from a squirt bottle, then flip the tube to discard the ethanol solution. Repeat the 70% ethanol rinse 2 times. Place the tube upside down on a clean paper towel for 5-10 min to air-dry the DNA pellet. Add 100 ul of TE buffer or deionized water to the DNA pellet, pipette or vortex vigorously to suspend the DNA. Centrifuge at 12,000 xg for 10 min to pellet any insoluble material, which contains residual PCR inhibitors, and transfer the clear DNA solution to a new tube.

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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