• Isolate all sizes of circulating and exosomal RNA, including microRNA
  • Versatile urine input ranging from 10 mL to 30 mL
  • 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 a flexible elution volume ranging from 50 µL to 100 µL
  • Purify high-quality RNA in 40 - 45 minutes

ItemsCat. #Size
Urine Cell-Free Circulating RNA Purification Maxi Kit 57100 10 preps

Urine Cell-Free Circulating RNA Purification Maxi Kit

This kit provides 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 from 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 all components for the purification are provided in one convenient and fast kit for the easy processing of small input volumes of bodily fluids. For a schematic workflow of the protocol click here.

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)

Protocol

Workflow

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 (15-25°C) for up to 2 years without showing any reduction in performance. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.


Kit Components
Component Cat. 57100 (10 preps)
Binding Solution K 1 x 75 mL, 1 x 25 mL
Lysis Buffer A 20 mL
Wash Solution A 18 mL
Elution Solution A 6 mL
Mini Spin Columns 10
Maxi Spin Columns 10
Collection Tubes 10
Elution Tubes (1.7 mL) 10
Product Insert 1

Product Information Sheets: