Introduction
The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA and miRNA from cell and tissue without MagZol reagent |
| Applications | RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Cell, Animal tissue, Plant tissue |
| Sample amount | Cells: ≤ 5 x 10^6, Animal tissue:<10mg |
| Elution volume | ≥30μl |
| Time per run | ≤40 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
Principle
Biological samples are first lysed and homogenized in a highly denaturing guanidine isothiocyanate-containing buffer, which immediately inactivates DNases and RNases to ensure isolation of intact DNA and RNA. The lysate is then passed through a Mini spin column. This column, in combination with the high-salt buffer, allows selective and efficient binding of genomic DNA. Flow-through from the column is digested by Proteinase K in the presence of ethanol. This optimized digestion, together with the subsequent addition of further ethanol, allows appropriate binding of total RNA, including miRNA, to the column. Contaminants are efficiently washed away and high-quality RNA is eluted.
Advantages
- High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
- Fast – several samples can be extracted in 40 minutes
- High applicability – samples including animals, plants, bacteria, cells, etc.
- High concentration – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity
- Safe – no phenol chloroform extraction and no use of Trizol reagent
Kit Contents
| Contents | R431102 | R431103 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Mini Columns | 100 | 2 x 250 |
| 2ml Collection Tubes | 100 | 2 x 250 |
| Proteinase K | 48 mg | 240 mg |
| Protease Dissolve Buffer | 5 ml | 15 ml |
| Buffer RLC | 40 ml | 200 ml |
| Buffer RWC | 20 ml | 80 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 60 ml |
Storage and Stability
Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
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