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Stool Total RNA and Genomic DNA Isolation Kit Add to Cart

Cat#: 45600-NB
Quantity: 50 preps
Price: 473 €
Supplier: Norgen
Shipping: RT
User Manual  

A convenient and rapid method to isolate total DNA and RNA from fresh or frozen stool samples

• Simultaneous isolation of both host and microbial DNA and RNA (Universal protocol)
• Eliminates PCR inhibitors including humic acid
• High quality total RNA and DNA for sensitive downstream applications
• Fast and easy processing - Rapid isolation of high-quality, ready-to-use total RNA and DNA in 30 minutes.
• No organic extraction or alcohol precipitation - Phenol is not required for the isolation
• High yields - Consistent, high yields of inhibitor-free RNA and DNA

Norgen’s Stool Nucleic Acid Isolation kit provides a convenient and rapid method to isolate total DNA and RNA from fresh or frozen stool samples. The universal protocol conveniently allows for the isolation of total genomic DNA and total RNA from all the various microorganisms and host cells found in the stool sample simultaneously. The kit removes all traces of humic acid using the provided Bead Tubes and a combination of chemical and physical homogenization and lysis, without the use of phenol-chloroform extractions.

Purification is based on spin column chromatography using Norgen´s proprietary resin as the separation matrix. Briefly, the stool sample is lysed using a combination of Lysis Solution and the provided Bead Tubes. The released RNA and DNA are then bound to Norgen´s column in the presence of Binding Solution and ethanol (BIND). Under these conditions only the RNA and DNA will bind to Norgen´s resin while most of the contaminatns and cellular proteinaceous components are removed in the flowthrough. The bound RNA and DNA are then washed to remove any remaining impurities (WASH). Lastly, the purified inhibiitor-free total RNA and DNA are eluted into the provided Elution Buffer or water (ELUTE). Please see procedure flowchart below.

Workflow
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The purified DNA and RNA are of the highest quality and are fully compatible with downstream PCR applications, as all humic acid substances and PCR inhibitors are removed during the isolation.


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Total RNA Profile from Different Stool Samples. Total RNA and DNA was isolated from 6 different 200 mg human stool samples using Norgen´s Stool Nucleic Acid Isolation Kit. For analysis, 7.5 µL from each 75 µL elution was loaded on 1.2 % MOPS agrarose gel. All six sample showed a good RNA integrity and total RNA profile that includes large and small RNA.


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Detection of Bacterial RNA from Stool Samples. Real-time one step RT-PCR analysis (SYBR Green) was carried out to detect the bacterial 16s rRNA expression from six stool samples isolated using Norgen’s Stool Nucleic Acid Isolation Kit. The RNA template was treated with DNase I and concentrated with Norgen’s RNA Clean-Up and Concentration Kit (Cat# 23600, 43200). No PCR inhibition was observed from 300 ng of total RNA in 20 µL PCR reaction. Black diamond: negative control, Blue diamond: six stool RNA samples.


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Isolation of Genomic DNA from Human Stool Samples. Total RNA and DNA was isolated from 6 different 200 mg human stool samples using Norgen´s Stool Nucleic Acid Isolation Kit. For analysis, 10 µL of each 75 µL elution was run on a 1.2% 1xTAE agarose gel. High quality DNA was isolated from all six stool samples. M: Norgen’s HighRanger 1kb DNA ladder (Cat. 11900).


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Detection of Human DNA from Stool Samples. Total RNA and DNA were isolated from two different stool samples using Norgen’s Stool Nucleic Acid Isolation Kit. Two microliters of each elution was used in 20 µL of Norgen’s 2X PCR Mastermix spiked with SYBR green (Bio-Rad), using 0.3 µM of primers against GAPDH. As can be seen in the amplification plot, all four samples were successfully amplified, indicating that PCR inhibitors were removed and the kit isolated high quality of DNA from stool. PCR quantification data (A) and melting curve analysis date (B) are shown.


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