Norgen’s Cell Culture Media Exosome Purification and RNA Isolation Kits constitute an all-in-one system for the purification of exosomes and the subsequent isolation of exosomal RNA from different cell culture media sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. The kit is 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 in that 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. 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.
Cell Culture Media Exosome Purification and RNA Isolation Mini Kit
For sample input volumes ranging from 5 mL to 10 mL.
Cell Culture Media Exosome Purification and RNA Isolation Midi Kit
For sample input volumes ranging from 10 mL to 20 mL.
Cell Culture Media Exosome Purification and RNA Isolation Maxi Kit
For sample input volumes ranging from 20 mL to 35 mL.
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| Kit Specifications | |
| Sample Type | Cell-Free Cell Culture Media |
| Sample Volume Range | 10 mL to 20 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 4 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. This kit is stable for 2 years from the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60ºC if any salt precipitation is observed.
| Component | Cat. 60700 (50 preps) | Cat. 60800 (25 preps) | Cat. 60900 (15 preps) |
|---|---|---|---|
| Slurry E | 12.5 mL | 12.5 mL | 12.5 mL |
| ExoC Buffer | 1.5 mL | 1.5 mL | 1.5 mL |
| ExoR Buffer | 12 mL | 12 mL | 12 mL |
| Lysis Buffer A | 20 mL | 20 mL | 20 mL |
| Lysis Additive B | 2 mL | 2 mL | 2 mL |
| Wash Solution A | 18 mL | 18 mL | 18 mL |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Mini Filter Spin Column | 50 | 25 | 15 |
| Mini Spin Columns | 50 | 25 | 15 |
| Collection Tubes | 50 | 25 | 15 |
| Elution tubes (1.7 mL) | 100 | 50 | 30 |
| Product Insert | 1 | 1 | 1 |
MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.
This method is suitable for small quantities of tissue (<100mg) and cells (<5 X106), and large quantities of tissue (up to 1g) and cells (<108), of human, animal, plant, or bacterial origin. The simplicity of the MagZol Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in one hour. Total RNA prepared in this manner can be used for Northern blot analysis, dot blot hybridization, poly(A) + selection, in vitro translation, RNase protection assay, and molecular cloning. For use in amplification by thermal cycling, treatment of the isolated RNA with RNase-free DNase I is recommended when the two amplimers lie within a single exon.
Specifications
| Features | Specifications |
| Main Functions | RNA isolation solvent (substitution for Trizol/Qiazol reagent) |
| Applications | RT-PCR, Northern hybridization, poly (a) enrichment, etc. |
| Purification technology | Acid phenol guanidine isothiocyanate |
| Process method | Manual (centrifugation) |
| Sample type | Animal tissue, plant tissue, adherent cells, suspended cells, bacteria, trace fungi, etc. |
| Sample amount | Flexible |
| Elution volume | Variation with sample size |
| Time per run | Variation with sample size |
Storage and Stability
MagZol Reagent should be stored at 2-8°C upon arrival and is stable for at least 24 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance.
Experiment Data
MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.
