Norgen’s Urine (Exfoliated Cell) RNA Purification Kit provides a rapid method for the isolation and purification of total RNA from exfoliated cells that have been shed into the urine from the urinary tract. The total RNA is preferentially purified from other cellular components such as proteins without the use of phenol or chloroform. RNA biomarkers from exfoliated cells can be used as non-invasive tools for a number of diagnostic and research applications including the diagnosis and monitoring of bladder, kidney, or other urinary-tract cancers. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other cellular components such as proteins, as well as from the contaminating species found in urine such as glucose and salts, without the use of phenol or chloroform. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. The process involves first lysing the exfoliated cells with the provided Lysis Solution. Ethanol is then added to the lysate, and the solution is loaded onto a spin-column (BIND). Under these conditions only the RNA will bind to Norgen’s resin while the contaminating proteins will be removed in the flowthrough or retained on the top of the resin. The bound RNA is then washed with the provided wash buffer in order to remove any remaining impurities (WASH). Lastly, the purified total RNA is eluted into 20 - 50 µL of the provided Elution Buffer or water (ELUTE). Please see procedure flowchart to the right.
FEATURES AND BENEFITS
- Fast processing -Rapid spin-column format allows for the processing of multiple samples in 20 minutes.
- Isolate diversity of RNA species - All RNA species can be isolated, from large mRNA and ribosomal RNA down to microRNA and small interfering RNA.
- Isolate RNA from small samples - RNA can be isolated and detected from as little as 100 exfoliated cells.
- No phenol:chloroform extractions - Total exfoliated cell RNA is isolated without the use of harmful chemicals such as phenol or chloroform.
- Recovered RNA is suitable for downstream applications - Purified RNA is fully compatible with real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.