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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.   

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.


Streamline fecal sample collection and DNA/RNA extraction
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.


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







Brief Protocol 


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.).






AquaStool human fecal DNA/RNA extraction protocol


1. Extract the DNA/RNA

Add 50 mg of human fecal sample to a 1.5-ml microfuge tube preloaded with 0.5 ml AquaStool solution and 100 ug of sand. Vortex the tube upside down at top speed for a few minutes (use a multichannel bead beater, if available). Add 250 ul of AquaRemove (#1208, order separately and dilute it with an equal volume of isopropanol before use) to the crude lysate. Vortex to mix well and centrifuge at 14,000 xg for 5 min to pellet the fecal debris.

2. Precipitate the DNA/RNA

Transfer 0.6 ml cleared lysate to a 1.5-ml tube and add 0.8 volume (0.48 ml) isopropanol. Vortex and centrifuge at 14,000 xg for 5 min to pellet the DNA/RNA. Decant to discard the supernatant. Rinse the pellet by filling the tube with 70% ethanol from a squirt bottle (be sure to rinse the entire interior of the tube, including the cap and the mouth of the tube), and decant to discard the ethanol solution. Repeat the ethanol rinse 2 times. Tap the tube on a clean paper towel to remove residual ethanol and air-dry the pellet for 4-5 min.

3. Solubilize the DNA/RNA

Add 0.4 ml deionized water to the pellet, Pipet and vortex to fully disperse the pellet. Incubate at 22 C for 15 min and then centrifuge at 14,000 xg for 5 min to pellet any insoluble. Transfer the clear DNA/RNA solution to a new tube. Store the DNA/RNA solution at 20 or -80 C.

 

 

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.

Note:    Order AquaRemove (# 1208, 8 ml for $49) separately for human fecal DNA/RNA purification. Dilute 8 ml AquaRemove with 8 ml isopropanol before use.

  

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MultiTarget Pharmaceuticals, LLC - All rights reserved