These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different urine sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits since they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA 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.
Urine Exosome and Free-Circulating RNA Isolation Mini Kit
For sample volumes ranging from 250 µL to 1 mL.
Urine Exosome and Free-Circulating RNA Isolation Midi Kit
For sample volumes ranging from 2 mL to 10 mL.
Urine Exosome and Free-Circulating RNA Isolation Maxi Kit
For sample volumes ranging from 11 mL to 30 mL.
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| Kit Specifications | |
| Minimum Urine Input | 250 µL |
| Maximum Urine Input | 1 mL |
| Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
| Elution Volume | 50 – 100 µL |
| Time to Complete 10 Purifications | 35 – 40 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 4 of the product insert for the average yields and the common RNA quantification methods.
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. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.
| Component | Cat. 59200 (50 preps) | Cat. 59300 (25 preps) | Cat. 59400 (15 preps) |
|---|---|---|---|
| Slurry E | 12.5 mL | 12.5 mL | 12.5 mL |
| ExoC Buffer | 8 mL | 30 mL | 50 mL |
| ExoR Buffer | 12 mL | 12 mL | 12 mL |
| Lysis Buffer A | 2 x 20 mL | 2 x 20 mL | 2 x 20 mL |
| Lysis Additive B | 2 mL | 2 mL | 2 mL |
| Wash Solution A | 2 x 18 mL | 18 mL | 18 mL |
| Elution Solution A | 2 x 6 mL | 6 mL | 6 mL |
| Mini Filter Spin Columns | 50 | 25 | 15 |
| Mini Spin Columns | 100 | 50 | 30 |
| Collection Tubes | 100 | 50 | 30 |
| Elution Tubes (1.7 mL) | 100 | 50 | 30 |
| Product Insert | 1 | 1 | 1 |
Norgen’s 2X One-Step RT-PCR Master Mix is a ready-to-use solution that contains components required for RT-PCR amplification of RNA templates. The mix includes M-MuLV reverse transcriptase, Taq DNA polymerase, dNTPs, reaction buffer, MgCl2, KCl, and a PCR enhancer/stabilizer. The user needs only to add the template, the primer set and water to the Master Mix to set up the RT-PCR reaction. This convenient 2X One-Step RT-PCR Master Mix reduces the time required to set up PCR reactions and reduces the possibility of contamination, particularly when preparing large numbers of reactions. The optimized master mix allows for robust amplification of RNA templates with high yields of PCR products.
Taq DNA Polymerase is a highly thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a very low 5´→ 3´ exonuclease activity. The source of Taq included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned Taq DNA Polymerase gene from Thermus aquaticus YT-1. M-MuLV Reverse Transcriptase is an RNA-directed DNA polymerase that can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. The source of the Reverse Transcriptase included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned reverse transcriptase gene from M-MuLV.
Norgen’s 2X One-Step RT-PCR Master Mix is available in 3 convenient sizes:
Cat # 28113 – 100 reactions (sufficient for 100 reactions x 20 µL reaction volume)
Cat # 28114 – 200 reactions (sufficient for 200 reactions x 20 µL reaction volume)
Cat # 28115 – 500 reactions (sufficient for 500 reactions x 20 µL reaction volume)
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Reagents supplied – Cat # 28114
Storage Conditions and Product Stability
2X One-Step RT-PCR Master Mix should be stored at -20°C. For everyday use an aliquot can be stored at 4°C for up to 3 months. Repeated freeze-thaw cycles are not recommended. When stored at the proper temperature this reagent is stable for at least 1 year.
K-AMYLSD
SKU: 700004263
160 assays (manual) / 640 assays (auto-analyser)