Norgen’s Total RNA Purification Maxi Kit provides a rapid method for the isolation and purification of total RNA from cultured animal cells, tissue samples, blood, bacteria, yeast, fungi, plants, and viruses. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other cellular components such as proteins, without the use of phenol or chloroform. The purified RNA is of the highest integrity, and 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.
Norgen’s Purification Technology
Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. The RNA is preferentially purified from other cellular components such as proteins without the use of phenol or chloroform. The process involves first lysing the cells or tissue of interest with the provided Buffer RL (please see the flow chart on page 4). Ethanol is then added to the lysate, and the solution is loaded onto a maxi spin column. Norgen’s resin binds RNA in a manner that depends on ionic concentrations. Thus only the RNA will bind to the column, while the contaminating proteins will be removed in the flowthrough or retained on the top of the resin. The bound RNA is then washed with the provided Wash Solution A in order to remove any remaining impurities, and the purified total RNA is eluted with the Elution Solution A. The purified RNA is of the highest integrity, and can be used in a number of downstream applications.
Figure 1 / 2
Click for expanded view
| Kit Specifications | |
| Maximum Column Binding Capacity | 1.5 mg |
| Maximum Column Loading Volume | 20 mL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Maximum Amount of Starting Material | |
| Liver Tissue | 100 mg |
| Heart Tissue | 50 mg |
| Kidney Tissue | 100 mg |
| Brain Tissue | 250 mg |
| Lung Tissue | 100 mg |
| Whole Blood | 2 – 10 mL* |
| Bacteria | 2.5 x 10 10 cells |
| Yeast | 2 x 108 cells |
| Fungi | 1 g |
| Plant Tissues | 1 g |
| Time to Complete 4 Purifications | 40 minutes |
| Average Yield: HeLa Cells (5 x 107 cells) E. coli (2.5 x 1010 cells) | ~750 μg ~1.5 mg |
*For isolating total RNA from purified leukocytes
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. 26800 (8 preps) |
|---|---|
| Buffer RL | 2 x 40 mL |
| Wash Solution A | 4 x 38 mL |
| Elution Solution A | 3 x 20 mL |
| RNA Maxi Spin Columns (with collection tubes) | 8 |
| Elution Tubes (50 mL) | 8 |
| Product Insert | 1 |
PEG3-(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane)-(Amino-Tri-(carboxyethoxymethyl)-methane) is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The terminal carboxylic acid groups can react with primary amino groups in the presence of activators (e.g. HATU) to form a stable amide bond.
PEG3-(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane)-(Amino-Tri-(carboxyethoxymethyl)-methane) is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The terminal carboxylic acid groups can react with primary amino groups in the presence of activators (e.g. HATU) to form a stable amide bond.
