This kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. The kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species without any requirement for phenol. Included in the kit are sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations. Ten samples can be processed in approximately 45 minutes. This kit is also available in a 100 prep size.
Background
In certain circumstances it is desirable to be able to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA.
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Kit Specifications | |
Maximum Column Binding Capacity | Up to 50 µg RNA |
Maximum Column Loading Volume | 650 µL |
Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
Time to Complete 10 Purifications | 45 minutes |
RNA Yield HeLa (1 x 106) – Cytoplasmic RNA HeLa (1 x 106) – Nuclear RNA | 15 µg ≤ 3.5 µg |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Component | Cat. 21000 (50 preps) | Cat. 37400 (100 preps) |
---|---|---|
Lysis Buffer J | 20 mL | 2 x 20 mL |
Buffer SK | 40 mL | 2 x 40 mL |
Wash Solution A | 38 mL | 2 x 38 mL |
Elution Buffer E | 6 mL | 2 x 6 mL |
Mini Spin Columns | 50 | 100 |
Collection Tubes | 50 | 100 |
Elution Tubes (1.7 mL) | 50 | 100 |
Product Insert | 1 | 1 |
ProbeSure Master Mix combines class-leading performance with the most competitive prices in the market. ProbeSure Master Mix has provided users with accurate, robust, and consistent performance for many years. It has been carefully optimised to work over a wide range of reaction volumes and sample template qualities.
ProbeSure Master Mix is supplied at 2x concentration for convenience and is supplied with the ROX normalising dye at either high level (500 nM final concentration), low level (25 nM final concentration) or without ROX.
Introduction
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.
The series of nucleic acid columns produced by Magen Biotech are based on carefully selected imported glass fiber membranes (GF/B, GF/D, GF/F). Columns production processes such as polypropylene injection molding materials, injection molding process, and downstream membrane packing and compression rings are strictly controlled. This is to ensure that the column has extremely high adsorption capacity and long-term stability. Compared with conventional products on the market, Magen’s columns are with varieties, and binding rate will not change when stored at room temperature for 4 years.
Specifications
Features | Specifications |
Recommended application | Plasmid Maxi preparation |
Preservation conditions | Room temperature |
Stability | Up to 4 years |
Filter membrane | High quality glass fiber filter GF/B, 8 layers |
Membrane aperture | 1.0 μm |
Maximum binding yield of plasmid | 1 mg |
Maximum yield of alcohol mediated binding | 5 mg |
Plasmid yield | Up to 1mg |
Single liquid carrying capacity of column | 20 ml |
Minimum elution volume | 800 μl |
Withstand centrifugal force | 3,000-5,000 x g |
Centrifuge | Low speed centrifuge for 50ml centrifuge tubes, >3000 x g, swing-out Rotor |
Adsorption Mechanism
Based on the negatively charged DNA skeleton, it has a high affinity for positively charged glass fibers. In high salt and ethanol solutions, DNA/RNA binds to glass fiber and interacts with hydrophilic matrix on silica through hydrogen bond. DNA/RNA is tightly bound. All pollutants can be removed by washing solution. At high salt concentration, nucleic acids selectively bind to silicagel membrane, while other pollutants, mainly proteins, are removed by membrane washing.
Ordering information
CAT.No. | Product Name | Package |
C13123 | HiPure DNA Maxi Column III (8 x GF/B)with 50ml Collection Tubes | 100/Bag |
Item No. | Product Name | Membrane type/number of layers | Collection tubes | Plasmid DNA binding capacity (Physical adsorption) | gDNA/RNA binding capacity (Alcohol-mediated adsorption) | Minimum Elution volume | Liquid volume capacity |
C13010 | HiPure DNA Nano Column | 2 layers GF/F | 2ml without cap | 5μg | 20μg | 10μl | 700μl |
C13011 | HiPure DNA Micro Column | 3 layers GF/F | 2ml without cap | 10μg | 50μg | 15μl | 700μl |
C13100 | HiPure DNA Mini Column I | 2 layers GF/B | 2ml without cap | 15μg | 100μg | 30μl | 700μl |
C13110 | HiPure DNA Mini Column II | 4 layers GF/B | 2ml without cap | 35μg | 200μg | 50μl | 800μl |
C13111 | HiPure RNA Mini Column | 3 layers GF/B | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
C13112 | HiPure Viral Mini Column | 3 layers GF/F | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
C13113 | HiPure CFDNA Mini Column | 3 layers GF/F,1 layer GF/B | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
C13120 | HiPure DNA Midi Column | 4 layers GF/B | 15ml Centrifuge tube | 125μg | 1mg | 500μl | 4ml |
C13121 | HiPure DNA Midi Column III | 8 layers GF/B | 15ml Centrifuge tube | 250μg | 1mg | 500μl | 4ml |
C13122 | HiPure DNA Maxi Column | 4 layers GF/B | 50ml Centrifuge tube | 500μg | 5mg | 1000μl | 20ml |
C13123 | HiPure DNA Maxi Column III | 8 layers GF/B | 50ml Centrifuge tube | 1mg | 5mg | 1000μl | 20ml |
C13124 | HiPure DNA Maxi Column C | 8 layers GF/B | 50ml high speed Centrifuge tube | 1mg | 5mg | 700μl | 12ml |
C13130 | HiPure DNA Plate | 2 layers GF/F | 1.6ml Plate | 30μg | 100μg | 80μl | 900μl |
C13131 | HiPure gDNA Plate | 2 layers GF/B | 1.6ml Plate | 30μg | 100μg | 80μl | 900μl |
Note: GF/B pore size is for 1.0μM glass fiber membrane; GF/F pore size is for 0.7μm glass fiber membrane.
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.
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