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Urine Cell-Free Circulating RNA Purification Kits

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Product Info

Overview

Isolate all sizes of circulating and exosomal RNA, including microRNA
Versatile urine input ranges
No phenol extractions
No carrier RNA
Bind and elute all RNA irrespective of size or GC content, without bias
Concentrate circulating RNA and exosomal RNA into flexible elution volumes
Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

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

Details

Supporting Data

Figure 1 / 9

Figure 1. Mini Kit Purification of cell-free circulating RNA and exosomal RNA from different urine volumes.

Figure 2. Mini Kit Linearity of RNA purified from increasing urine volumes.

Figure 3. Mini Kit Determination of the amount of inhibition present in urine cell-free circulating RNA samples when detecting the human miR-21.

Figure 4. Midi Kit Purification of cell-free circulating RNA and exosomal RNA from different urine volumes.

Figure 5. Midi Kit Linearity of RNA purified from increasing urine volumes

Figure 6. Midi Kit Determination of the amount of inhibition present in urine cell-free circulating RNA samples when detecting the human miR-21

Figure 7. Maxi Kit Purification of cell-free Circulating RNA and exosomal RNA from different urine volumes

Figure 8. Maxi Kit Linearity of RNA purified from increasing urine

Figure 9. Maxi Kit Determination of the amount of inhibition present in urine cell-free circulating RNA samples when detecting the human miR-21

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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