These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating RNA, including exosomal RNA as well as viral RNA from fresh, preserved or frozen urine samples. All components for the purification are provided in one convenient and fast kit for the easy processing of small input volumes of bodily fluids. The purified urine RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.
Background
Recent evidence indicates that cell-free circulating RNA (cf-RNA) including exosomal RNA in urine contains valuable information for the discovery of biomarkers that can help for the early detection of certain cancer types and for monitoring the disease status as well as for the detection of any infectious pathogens. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. 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. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. The advantage for using urine as a source for cancer biomarkers is that it can be acquired in large quantities without using invasive procedures. In addition, repeated sampling from the same individual is applicable, which facilitates longitudinal studies. There are many advantages favouring the use of urinary nucleic acid for cancer biomarker discovery over blood, tissue samples or other bodily fluids, including: (1) urine is non-infectious for HIV and less infectious for many other pathogens; (2) the profile of urinary nucleic acid is similar to that in plasma or serum but with a lower concentration; (3) Nucleic acid purification from urine is technically much easier because of its low protein concentration (1000-fold lower than blood).
Urine Cell-Free Circulating RNA Purification Mini Kit
For sample volumes ranging from 250 µL to 2 mL.
Urine Cell-Free Circulating RNA Purification Midi Kit
For sample volumes ranging from 2 mL to 10 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
Urine Cell-Free Circulating RNA Purification Maxi Kit
For sample volumes ranging from 10 mL to 30 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL
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| Kit Specifications | |
| Minimum Urine Input | 10 mL |
| Maximum Urine Input | 30 mL |
| Size of RNA Purified | All sizes including miRNA and small RNA (< 200 nt) |
| Elution Volume | 50-100 μL |
| Time to Complete 10 Purifications | 40-45 minutes |
| Average Yields | Variable depending on specimen |
*Please check page 6 of the product insert for average yields and common RNA quantification methods.
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. These kits are stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
| Component | Cat. 56900 (50 preps) | Cat. 57000 (20 preps) | Cat. 57100 (10 preps) |
|---|---|---|---|
| Binding Solution K | 25 mL | 75 mL | 1 x 75 mL 1 x 25 mL |
| Lysis Buffer A | 30 mL | 20 mL | 20 mL |
| Wash Solution A | 18 mL | 18 mL | 18 mL |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Mini Spin Columns | 50 | 20 | 10 |
| Midi Spin Columns | – | 20 | – |
| Maxi Spin Columns | – | – | 10 |
| Collection Tubes | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
Biotin-PEG8-alkyne is a PEG linker containing a biotin group and a terminal alkyne. Biotin is useful for affinity-based applications such as pull-down assays while terminal alkynes are used in copper (I) click chemistry with azide groups on a target molecule. The inclusion of a PEG linker in this molecule improves its solubility in water.
Biotin-PEG8-alkyne is a PEG linker containing a biotin group and a terminal alkyne. Biotin is useful for affinity-based applications such as pull-down assays while terminal alkynes are used in copper (I) click chemistry with azide groups on a target molecule. The inclusion of a PEG linker in this molecule improves its solubility in water.
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DBCO-NHCO-PEG4-NHS ester is a PEG linker containing NHS ester that is able to react specifically and efficiently with primary amines such as the side chain of lysine residues or aminosilane-coated surfaces at neutral or slightly basic condition to form a covalent bond. The hydrophilic PEG spacer arm improves water solubility and can also provide a long and flexible connection that minimizes steric hindrance during ligation. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.