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 | 11 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 the specimen |
*Please check page 5 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 shipping. 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 microScript microRNA cDNA Synthesis Kit is an all-in-one, ready-to-use product for the reverse transcription of microRNA from either Total RNA preparations or enriched microRNA preparations. The kit contains the 2x Reaction Mix and the microScript microRNA Enzyme Mix. The kit utilizes Norgen’s microScript Reverse Transcriptase, a mutant version of Moloney Murine Leukemia Virus (M-MuLV) Reverse Transcriptase. It has reduced RNase H activity and increased thermal stability.
The workflow of Norgen’s microScript microRNA cDNA Synthesis Kit involves a simple, single-tube set-up by the mixing of 2x Reaction Mix, Enzyme Mix and the RNA template. The reaction can then be carried out in a thermocycler. A poly (A) tail is first added to the RNA template, followed by cDNA synthesis using an adapter primer. In addition to the ease-of-use, the single-tube set-up provides superb consistency and sensitivity. The cDNA could be used in a PCR or qPCR amplification using a Universal PCR Reverse Primer and the forward primer that contains the sequence of the microRNA of interest. A single cDNA preparation could be used for PCR amplification of a number of different microRNAs. In addition, the cDNA preparation could be used for PCR or qPCR detection (using gene-specific forward and reverse primers) of mRNA or large RNA if total RNA preparation was the starting template. This could allow for parallel evaluation of expression level of microRNAs and microRNA-targets.
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Storage Conditions and Product Stability
Norgen’s microScript microRNA cDNA Synthesis Kit components should be stored at -20°C. These reagents should remain stable for at least 1 year in their unopened containers.
| Component | Cat. 54415 (12 rxns) | Cat. 54410 (50 rxns) |
|---|---|---|
| microScript microRNA Enzyme Mix | 12 μL | 50 μL |
| 2x Reaction Mix | 120 μL | 500 μL |
| Universal PCR Reverse Primer | 60 μL | 250 μL |
| Nuclease-Free Water | 1.25 mL | 2 x 1.25 mL |
| Product Insert | 1 | 1 |
* Optical system: equipped with infinite flat field optical system, the whole machine mildew prevention design and large field eyepiece, with beautiful appearance, clear imaging, broad vision, convenient operation and other characteristics, can be widely used in biology, medicine, industry, agriculture and other fields, is the ideal instrument for medical, teaching, scientific research and other units
* Observation head device: the cylinder can rotate 360 degrees, the displacement of the center of the mirror is 0.06mm; 30 tilt; the hinge type binocular observation (the magnification difference between left and right systems is 2.0%; the end position difference is 0.1mm at zero vision) and the third party connection port, the pupil distance adjustment range is 50-75mm, the fixed observation cylinder head is internal hexagon screw or select portable screw.
* Eyepiece configuration: random eyepiece WF10X / 22mm; the magnification accuracy of eyepiece shall not exceed ± 2%. There is a card slot on the eyepiece tube to lock the eyepiece to prevent falling. Binocular vision is adjustable, making binocular observation easier;
* Objective: Broadband coated ED-N195 pure copper infinite remote flat field achromatic objective; objective magnification accuracy not exceeding ± 5%; 4X, 10X, 40X (spring), 100X (spring, oil); objective error 10-40 Aim 0.06mm; 40-100 Aim 0.03mm
* Loading platform: triangular rail XY composite mechanical moving loading platform; 0.005 lateral to 5N horizontal force, corrosion resistance and wear resistance. Rectangular, size 190X140mm, stroke 6090mm, minimum read value 0.1 mm.
Parameter of microscope
| Specification configuration | TL36A | Binocular observation head, 45° tilt, 360 degrees can be rotated, pupil distance can be adjusted, adjustment range 50-75mm |
| TL36B | Three-eye observation head, 45° tilt, 360 degrees can be rotated, pupil distance can be adjusted, adjustment range 50-75mm. | |
| TL36M | Three-eye viewing head with 6.3 megapixel imaging system CMOS camera | |
| TL36P | Three-eye viewing head, equipped with 10.1 inch integrated flat panel display | |
| Eyepiece | Wide-angle eyepiece WF10X,field of view 22mm,optional WF16X,WF25X | |
| Converter | Quadruple Revolving converter | |
| 0bjective | Flat field achromatic objective at infinite distance,4X,10X,40X,100X(oil) | |
| Condenser | ABBE N.A.1.25Condenser with Iris Diaphragm &Filter | |
| Stage | Double Layer Mechanical Stage, Size1190X140mm, Moving Range : 60mmx90mm | |
| Focusing | 0.02mmCrude motion, micromotion and coaxial focal adjustment system with a minimum focal range of 0.02mm Coarse and Fine Focusing | |
| Light source | 3W LED,with Rechargeable Battery,Type C USB cable | |
| Attachments | Dust bag,Type C power cord | |
TL 36 series biological microscopes are equipped with infinite distance flat field achromatic objective and large field eyepiece, with the characteristics of beautiful appearance, clear imaging, broad vision, convenient operation and so on, can be widely used in biology, medicine, industry, agriculture and other fields, is the ideal instrument for medical, teaching, scientific research and other units