We have been using Norgen's using the Stool Nucleic Acid Collection and Preservation Tubes and Stool Total RNA Purification Kit
for many years for our projects focused on searching fecal biomarkers (small non-coding RNAs and bacterial species) to diagnose colorectal cancer non-invasively.
The challenge was to collect stool samples and preserve nucleic acids in a way that patients could collect samples at home and bring to the receiving hospital
without rush and preserving the samples. We successfully used the collection tubes for over 1000 samples in the frame of an European project and we were able to
extract either RNA and microbial DNA for metagenome analysis.
Barbara Pardini, Italian Institute for Genomic Medicine
In addition, the RNA purification kit proved also to be very efficient in the RNA extraction
also from leftover samples derived from the fecal immunochemical test (FIT) used in Italy to screen the general population for colorectal cancer.
Moreover, from these types of samples we could run small RNA sequencing from several hundreds of samples.
The products worked perfectly and helped us to engage new molecular approaches for implementing colorectal screening in a non-invasive way.
Alessio Naccarati, Italian Institute for Genomic Medicine
Stool Total RNA Purification Kit
A convenient and rapid method to purify total RNA from small amounts of stool samples.
For research use only and NOT intended for in vitro diagnostics.
Stool Total RNA Purification Kit (Cat. 49500)50 preps
Format: Spin Column
Norgen Biotek offers kits in different standardized formats. You can view and compare kit components in the Components Table in the Product Overview section. For more information on format types, please visit our Format Information page.
Simultaneous isolation of both host RNA and microbial RNA (universal protocol)
Isolate full diversity of RNA from large RNA down to small and microRNAs
Eliminates PCR inhibitors including humic acids
High quality RNA for sensitive downstream application
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
This kit provides a convenient and rapid method to purify total RNA from small amounts of stool samples. All types of stool samples can be processed with this kit, including animal fecal samples, manure and samples collected using Norgen's Stool Nucleic Acid Collection and Preservation Tubes (Cat. 45660). The kit removes all traces of humic acids using rapid and simple spin column procedures. Bead tubes are also provided for effective homogenization of stool. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA and small interfering RNA. Both host and microbial RNA is recovered. The protocol does not rely on the use of phenol or chloroform, thereby providing a user friendly procedure and allowing high-throughput analysis. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR and reverse transcription PCR for gene expression analysis. The procedure can be completed in approximately 30 minutes for 10 samples.
Figure 1. High yields of stool total RNA were purified from 200 mg of human stool using Norgen's Stool RNA Purification Kit and a leading competitor's kit, in triplicate. For analysis, 7.5 µL of each 75 µL elution was loaded on a 1.2 % 1x MOPS formaldehyde-agarose gel. Norgen's kit was found to have a higher yield of RNA, isolated from 200 mg of human infant stool.
Figure 2. Yield and Quality of Purified Stool RNA. Total RNA was isolated from 200 mg of human stool using Norgen's Stool RNA Purification Kit and a leading competitor kit. Comparisons were then made based on yield, and A260:A280/A260:A230 ratios measured using the NanoVue Plus™. In Panel A it can be seen that both kits isolated RNA with high A260:A280 ratios (all samples were found to be above 1.8 and below 2.2). In Panel B, Norgen's kit was found to isolate RNA with a high A260:A230 (with all samples once again falling in the 1.8-2.2 range). The competitor kit, however, was found to isolate RNA with extremely low A260:A230 ratios, with none of the samples displaying a A260:A230 ratio higher than 0.20. The results in Panel C are in agreement with the gel photo from Figure 1, and Norgen's kit was found to isolate higher amounts of RNA, with an average yield of 14.58 µg, compared to the competitors average of 12.26 µg.
Figure 3.Detection of Bacterial Stool RNA using 16S Primers. Total stool RNA was isolated from 200 mg human stool samples using Norgen's Stool RNA Purification Kit and a leading competitors kit. Five microliters of purified RNA was used in a 20 µL reverse-transcription reaction using Invitrogen's Superscript III system with 16S reverse primers. The cDNA generated was then used in a qPCR reaction involving Norgen's 2X PCR Mastermix spiked with SYBR green (Bio-Rad), using 0.3µM of primers against bacterial 16S. As can be seen in the amplification plot, Norgen's kit outperformed the leading competitor's kit by on average 1.5 Ct values. This indicates that Norgen isolated higher quality and yields of RNA from stool, that can be used in an array of downstream applications.