N-(Propargyl-PEG4-carbonyl)-N-bis(PEG1-acid) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The terminal carboxylic acids can react with primary amino groups in the presence of activators (e.g. HATU) to form a stable amide bond.
N-(Propargyl-PEG4-carbonyl)-N-bis(PEG1-acid) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The terminal carboxylic acids can react with primary amino groups in the presence of activators (e.g. HATU) to form a stable amide bond.
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 | 2 mL |
| Maximum Urine Input | 10 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 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 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 |
DBCO STP ester is a water-soluble reagent with a terminal DBCO group and a STP ester group. DBCO STP ester that can be used for the modification of peptides, antibodies, proteins, and other molecules containing the amine group. The STP esters are excellent alternatives to conventional N-hydroxysuccinimide (NHS) esters for coupling reactions in aqueous environments. The DBCO groups is commonly used for copper-free Click Chemistry reactions due to its strain promoted high energy. Reagent grade, for research use only.
DBCO STP ester is a water-soluble reagent with a terminal DBCO group and a STP ester group. DBCO STP ester that can be used for the modification of peptides, antibodies, proteins, and other molecules containing the amine group. The STP esters are excellent alternatives to conventional N-hydroxysuccinimide (NHS) esters for coupling reactions in aqueous environments. The DBCO groups is commonly used for copper-free Click Chemistry reactions due to its strain promoted high energy. Reagent grade, for research use only.
The HiPure Mini system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Plasmid Mini Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 70μg plasmid DNA from 5-15ml bacterial culture |
| Applications | Enzyme digestion, sequencing, PCR, cloning, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Conventional plasmid, plasmid less than 30KB |
| Sample amount | High copy plasmid: 1-5ml culture mediumLow copy plasmid : 5-10ml culture medium |
| Yield | 20-80µg |
| Elution volume | ≥75μl |
| Time per run | Complete 1-24 samples in 30 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 70µg |
The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
Kit Contents
| Contents | P100202 | P100203 |
| Purification Times | 50 Preps | 250 Preps |
| RNase A | 5 mg | 20 mg |
| Buffer P1 | 30 ml | 140 ml |
| Buffer P2 | 30 ml | 140 ml |
| Buffer P3 | 40 ml | 200 ml |
| Buffer PW1 | 30 ml | 140 ml |
| Buffer PW2* | 12 ml | 50 ml |
| Elution Buffer | 15 ml | 30 ml |
| HiPure DNA Mini Columns III | 50 | 250 ml |
| 2 ml Collection Tubes | 50 | 250 ml |
Storage and Stability
The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.
For any technical problems or customized products, please contact us.
F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here
The HiPure Mini system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Plasmid Mini Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.
