The Small RNA Library Prep Kit for Illumina consists of all the reagents and components required to generate small RNA libraries to be used for next-generation sequencing on an Illumina platform. All molecular reagents including adaptors, primers, enzyme mixes and buffers are provided. A purification module is also provided for rapid purification of nucleic acid products generated at various steps of the workflow. The purification module utilizes Norgen’s patent resin technology which enhances recovery of desired library intermediates or final products. The library prep workflow could be used for different forms of input including purified total RNA or enriched small RNA, as well as RNA from low content inputs such as plasma, serum and urine.
Workflow
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| Kit Specifications | |
| Number of preps | 24 |
| Number of indices provided | 12 (for 24 preps, indexes 1-24 or 25-48) |
| Time required to complete 6 libraries | As little as 5 hours |
| Input RNA required | As little as 50 pg |
Storage Conditions and Product Stability
Some components require storage at -20°C, 4°C or room temperature. See individual components and box labels for storage conditions.
| Step | Component | Cat. 64600 (24 preps) |
|---|---|---|
| 3′ AdaptorLigation to Template RNA | 3′ Adaptor | 30 µL |
| 3′ Adaptor Ligation Master Mix | 320 µL | |
| T4 RNA Ligase 2 (Truncated) | 35 µL | |
| 5′ Adaptor Ligation | 5′ Adaptor | 30 µL |
| 5′ Adaptor Ligation Master Mix | 320 µL | |
| T4 RNA Ligase 1 | 35 µL | |
| cDNA Synthesis from Ligated RNA Product | Reverse Primer | 30 µL |
| cDNA Synthesis Master Mix | 220 µL | |
| TruScript ReverseTranscriptase | 35 µL | |
| PCR Amplification | 2x NGS PCR Master Mix | 1.32 µL |
| PCR Reverse Primer | 81 µL | |
| Forward Index Primer | Included in Small RNA Library Prep Forward Index Primers (# 64640 or # 64610) | |
| Size Selection | NGS MW Ladder | 50 µL |
| NGS Control Ladder | 50 µL | |
| Loading Dye | 300 µL | |
| Nuclease-Free Water | 1.25 µL |
| Component | Cat. 63500 (75 preps) |
|---|---|
| Buffer RL | 40 mL |
| Wash Solution A | 38 mL (Reconstituted to 128 mL) |
| Elution Solution A | 6 mL |
| Columns | 75 |
| Gel Filtration Columns | 24 |
| Collection Tubes | 75 |
| Elution Tubes | 75 |
| Product Insert | 1 |
The Urine Exosome Purification Kits provide a fast, reliable and convenient method to purify and enrich for pure intact exosomes from different urine sample volumes ranging from 250 µL to 30 mL. These kits also allow for the purification of intact extracellular vesicles (EVs) from different urine sample volumes, and these EVs are ready for any downstream application. The purification is based on Norgen’s proprietary resin.
These kits provide a clear advantage over other available methods since they do not require any special instrumentation, ultracentrifugation, precipitation reagents or any protease treatments. More importantly, the purified exosomes will not be contaminated with any other RNA-binding proteins that may contaminate your exosomal RNA, which is essential if studying exosomal RNA gene expression.
NanoSight® Analysis
Exosomes enriched with Norgen’s Urine Exosome Purification Kits can be analyzed using NanoSight® for assessing the approximate exosome size range and concentration.
Exosomal RNA Analysis
Exosomal RNA can be isolated using Norgen’s Exosomal RNA Isolation Kit from exosomes enriched using Norgen’s Urine Exosome Purification Kits for gene expression analysis using RT-qPCR, microarray or NGS and for Biomarker discovery.
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| Kit Specifications | |
| Urine Input (Cat. 57700) | 250 μL – 1 mL |
| Urine Input (Cat. 57800) | 2 mL – 10 mL |
| Urine Input (Cat. 57900) | 11 mL – 30 mL |
| Size of Exosomes Purified | 40 nm – 150 nm |
| Elution Volume | Variable depending on the urine input volume |
| Time to Complete 10 Purifications | 15 – 30 minutes |
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.
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. 57700 (50 preps) | Cat. 57800 (25 preps) | Cat. 57900 (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 |
| Mini Filter Spin Columns inserted into 2 mL tubes | 50 | 25 | 15 |
| Product Insert | 1 | 1 | 1 |
Usages:
Beef extract powder is a purified extract of beef .Used with meat and casein peptones as a source of nutrients in culture media .
Technical specification:
Total nitrogen(TN)—————————≥12.0%
Amino nitrogen(AN) ————————≤2.7%
Chlorides————————————–＜6%
Moisture—————————————≤5.0%
PH———————————————–6-7
Storage: Keep container tightly closed, store in a cool, dry place, away from bright light.
Specifications: 500g/bottle; 10kg /bag
500g/bottle;10KG/bag