Catalog# : NB-22500
Price/Unit : $205.00
Unit : 20 preps
For the rapid purification of total RNA (including microRNA) from exfoliated cells in urine.
* Isolate and detect total RNA from as little as 1 mL of urine
* Provides high quality RNA for sensitive applications
* Rapid processing time from sampling to downstream testing
Bio-Synthesis'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 Bio-Synthesis'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 Bio-Synthesis'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.
APPLICATIONS
The purified RNA is of the highest quality and can be used in a number of downstream applications including:
* PCR
* Reverse Transcription PCR
* Northern blotting
* RNase protection
* Primer extension
* Expression array assays
Figure 1. Isolation and Detection of RNA from the Exfoliated Cells in 50 mL of Urine. RNA was isolated from the exfoliated cells in a 50 mL urine sample obtained from a healthy male without any spiking using Bio-Synthesis's Urine (Exfoliated Cell) RNA Purification Kit. The bind, wash and elute procedure was followed, and the purified RNA was recovered in 20 µL of the Elution Buffer. A dilution series of HeLa cells with known concentration (from 1 cell per mL to 1,000 cells per mL) was isolated in parallel as standards. Seven microliters of the 20 µL eluted RNA was then subjected to a 20 µL reverse transcription. One microliter of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the ribosomal S14 transcript. The red lines in the PCR baseline graph above correspond to the HeLa RNA standards, while the blue lines correspond to the successful PCR results when RNA isolated from the exfoliated cells in 50 mL of non-spiked urine was used as the template.Urine (Exfoliated Cell) RNA Purification Kit Components
1. Lysis Solution
2. Wash Solution
3. Elution Buffer
4. Micro Spin Columns
5. Collection Tubes
6. Elution tubes
7. Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.
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