The Kit provides fast purification of high-quality RNA from plants, cell, tissues, and yeast 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 precipitation with isopropanol or LiCl. RNA purified using the RaPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from <150 mg simple plant sample (including plant sample containing starch) without chloroform |
| Applications | RT-PCR, qPCR, Northern hybridization, second generation sequencing, nucleic acid protection, in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology, DNA filtration technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Economic crops |
| Sample amount | ≤150 mg |
| Elution volume | ≥30μl |
| Time per run | ≤25 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
Principle
The Kit isolates total RNA from up to 150mg plant tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 min. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanolis added to the flow-through, and the sample is applied to an RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit.
Advantages
- Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
- High quality – high purity total RNA can be directly used in various sensitive downstream applications
- Fast – several samples can be extracted in 25 minutes by column method
- Safe – no phenol chloroform extraction required
- Sensitive – RNA can be recovered at the level of PG
- Broad spectrum – various types of plant samples can be processed by diversity of operating procedures
Kit Contents
| Contents | R415102 | D415103 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure DNA Mini Column II | 50 | 250 |
| HiPure RNA Mini Columns | 50 | 250 |
| 2ml Collection Tubes | 100 | 2 x 250 |
| Buffer RLC | 50 ml | 200 ml |
| Buffer PRC1 | 50 ml | 200 ml |
| Buffer RW1 | 50 ml | 200 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 30 ml |
Storage and Stability
The Kit 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 inthe Buffer RLC/PRC1. Dissolve by warming buffer to 37°C.
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