Bio-Synthesis Urine Bacteria RNA Purification Kit is designed for the rapid preparation of bacterial RNA from urine samples. Bacterial RNA can be isolated from both human urine samples and urine samples from animals in order to study the levels and types of bacteria that are present, as well as to study the stage of bacterial pathogenesis through the use of RNA biomarkers.
The kit allows for the isolation of RNA from both Gram negative and Gram positive bacteria, including E. coli, Proteus spp., Klebsiella spp., Enterobacter spp., Serratia spp., Pseudomonas spp, Clostridial ssp. and Leptospirosis spp., as well as Chlamydia trachomatis and Neisseria gonorrhoeae.
The Urine Bacteria RNA Purification 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 proprietary resin as the separation matrix. The process involves first pelleting the urine bacterial cells, and then lysing the cells with the provided Lysis Solution.
Binding Solution and ethanol are added to the lysate, then it is loaded onto a spin-column (BIND). Under these conditions only the RNA will bind to Bio-Synthesis 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 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). Refer the procedure flowchart given below.
Features and Benefits
- Fast processing -Rapid spin-column format allows for the processing of multiple samples in 30 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 all types of bacteria found in urine -Bacterial RNA can be isolated from both Gram negative and Gram positive bacteria found in urine using the kit.
- No phenol:chloroform extractions - Urine bacterial 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 RT-PCR, quantitative RT-PCR and Northern Blot analysis.
The purified RNA is of the highest quality and can be used in a number of downstream applications including:
- Real-time PCR
- Reverse transcription PCR
- Northern blotting
- RNase protection
- Primer extension
- Expression array assays