This product is designed for purification of high-molecular-weight genomic or mitochondrial DNA from a variety of tissue and culture cells. The convenient, scalable purification procedure removes contaminants and enzyme inhibitors such as proteins and divalent cation, and purified DNA is ready for immediate use in sensitive downstream applications or for archiving.
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from tissue using economic salt out method |
| Applications | PCR, enzyme digestion, Southern hybridization,etc. |
| Purification technology | Salting out method |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Animal tissue |
| Sample amount | Unlimited |
| Elution volume | ≥100μl |
| Time per run | Variation with sample amount |
Principles
Cells are lysed with ananionic detergent in the presence of a DNA stabilizer. The DNA stabilizer limits the activity of intracellular DNases and also DNases found elsewhere in the environment. RNA is then removed by treatment with an RNA digesting enzyme. Other contaminants, such as proteins, are removed by salt precipitation. Finally, the genomic DNA is recovered by precipitation with alcohol and dissolved in Buffer TE. Purified DNA typically has an A260/A280 ratio between 1.7 and 1.9, and is up to 200 kb in size. The DNA can be safely stored at 2-8°C, -20°C, or -80°C.
| Contents | D331201 | D331202 | D331203 |
| Purification Times | 1 g | 5 g | 50 g |
| Proteinase K | 330 µl | 1.8 ml | 18 ml |
| Buffer WTL | 33 ml | 160 ml | 2 x 800 ml |
| Buffer PPS | 12 ml | 55 ml | 500 ml |
| RNase Solution | 330 µl | 1.8 ml | 18 ml |
| Buffer TE | 12 ml | 60 ml | 200 ml |
RNase Solution should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
This product is designed for purification of high-molecular-weight genomic or mitochondrial DNA from a variety of tissue and culture cells. The convenient, scalable purification procedure removes contaminants and enzyme inhibitors such as proteins and divalent cation, and purified DNA is ready for immediate use in sensitive downstream applications or for archiving.
The Kit provides fast purification of high-quality RNA from whole blood, cells and tissues using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. The purified RNA can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection and in vitro translation.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from 1-1.5ml whole blood |
| Applications | qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection |
| Purification method | Mini spin column |
| Purification technology | Silica technology, acidphenol / guanidine extraction technology (MagZol pretreatment technology) |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Fresh or frozen blood, mammalian blood |
| Sample amount | ≤1.5 ml whole blood |
| Yield | 2-100μg |
| Elution volume | ≥20μl |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100μg |
The Kit simplifies isolation of RNA from blood with a fast spin-column procedure. Red blood cells are selectively lysed and white cells collected by centrifugation. White cells are then lysed using highly denaturing conditions which immediately inactivate RNases. After homogenization using the DNA spin column, the sample is applied to the RNA column. Total RNA binds to the membrane and contaminants are washed away, leaving pure RNA to be eluted in 30–100µl RNase-free water (provided with the kit) for direct use in any downstream application.
Advantages
Kit Contents
| Contents | R416102 | R416103 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure DNA Mini Columns | 50 | 250 |
| HiPure RNA Mini Columns I | 50 | 250 |
| 2ml Collection Tubes | 100 | 500 |
| 10 x Buffer RBC | 50 ml | 3 x 100 ml |
| RTL Lysis Buffer | 50 ml | 250 ml |
| Buffer RW1 | 50 ml | 250 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 30 ml |
Storage and Stability
HiPure Blood RNA Mini Kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form in the RTL Lysis Buffer. Dissolve by warming buffer to 37°C.
The Kit provides fast purification of high-quality RNA from whole blood, cells and tissues using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. The purified RNA can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection and in vitro translation.
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 | 5 mL to 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 to 40 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 60°C 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 |