This kit provides a rapid spin column procedure for the isolation of exosomal RNA from urine samples. Users can simultaneously concentrate and isolate high quality exosomal RNA, including microRNA, for use in sensitive downstream assays such as RT-PCR, qRT-PCR, NGS, microarrays and more. The protocol can be completed in under 50 minutes. Urine volumes of 1 to 10 mL can be processed easily and rapidly. All sizes of RNA are recovered at an equal rate without the need for using hazardous chemicals like phenol.
Background
Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascite fluids, among other biological fluids. These vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which can inform about the cell type from which the exosomes are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNases they can be efficiently recovered from biological fluids, such as urine.
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| Kit Specifications | |
| Minimum Urine Input | 1 mL |
| Maximum Urine Input | 10 mL |
| Size of RNA Purified | Small exosomal RNA species |
| Time to Complete Purification | ~ 50 minutes |
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
| Component | Cat. 47200 (50 preps) |
|---|---|
| Slurry B1 | 18 mL |
| Binding Buffer A | 20 mL |
| Lysis Buffer A | 2 x 20 mL |
| Wash Solution A | 38 mL |
| Elution Solution A | 6 mL |
| Mini Filter Spin Columns | 50 |
| Collection Tubes | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
The BK virus is a member of the polyomavirus family. It has been suggested that this virus may be transmitted through respiratory fluids or urine, since infected individuals periodically excrete virus in the urine. BK viral infections are typically asymptomatic in healthy individuals, however very mild symptoms may appear including mild respiratory infections and fever. Infections with BK virus in immunocompromised or immunosupressed patients are much more severe and may involve renal dysfunction. In fact, in kidney transplant patients the immunosupressive drugs required for the transplant may allow the virus to replicate within the graft, resulting in a disease called BK virus nephropathy (BKVN). The JC virus is a type of human polyomavirus and is very common in the general population, infecting 70 to 90% of humans. Most people acquire JCV in childhood or adolescence. Typically the infection is subclinical and no of consequence in individuals with healthy immune systems. The initial site of infection may be the tonsils or the gastrointestinal tract, and the virus then remains latent in the gastrointestinal tract. JCV can also infect the tubular epithelial cells in the kidneys, where it continues to reproduce, shedding virus particles in the urine. Also, JCV can cross the blood-brain barrier into the central nervous system. JCV is known to cause the usually fatal progressive multifocal leukoencephalopathy (PML) by destroying oligodendrocytes in the brain in immunodeficient or immunosuppressed individuals. The JC and BK viruses are very similar, with their genomes sharing 75% homology. It is however important to differentiate between the viruses due to the differences in pathology and especially the invariably fatal outcome of PML which is only caused by the JC virus.
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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival.
| Component | Cat. TM39350 (100 preps) | Cat. TM39310 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| BKV/JCV Primer & Probe Mix | 280 μL | 280 μL |
| BKV/JCV Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
endo-BCN-PEG4-Biotin is a biotinylated PEG linker containing a BCN moiety for click chemistry and a PEG4 arm that imparts water solubility for the biotinylated molecule.
endo-BCN-PEG4-Biotin is a biotinylated PEG linker containing a BCN moiety for click chemistry and a PEG4 arm that imparts water solubility for the biotinylated molecule.
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