Norgen’s Stool Total RNA Purification 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 and manure. The kit removes all traces of humic acid using the provided Bead Tubes and a combination of chemical and physical homogenization and lysis. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA and small interfering RNA. The protocol does not rely on the use of phenol or chloroform, thereby providing a user friendly procedure and allowing high-throughput analysis on the lab bench.
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 is then bound to Norgen's column in the presence of ethanol (BIND). Under these conditions only the RNA will bind to Norgen's resin while most of the contaminating DNA and cellular proteinaceous components are removed in the flowthrough. The bound RNA is then washed to remove any remaining impurities (WASH). Lastly, the purified inhibiitor-free RNA is eluted into 75 µL of the provided Elution Buffer or water (ELUTE). Please see procedure flowchart to the right.
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.
FEATURES AND BENEFITS
- Fast and easy processing - Rapid isolation of high-quality, ready-to-use RNA in 30 minutes.
- No organic extraction or alcohol precipitation - Phenol is not required for the isolation
- Universal procedure - Simultaneous isolation of both host RNA and microbial RNA
- High yields - Consistent, high yields of inhibitor-free RNA
- Remove all inhibitors - Eliminates all PCR inhibitors including humic acid