Introduction
The HiPure Plant RNA Midi Kit provides fast purification of high-quality RNA from plants, cell, tissues, and yeast using silica-membrane spin columns with a binding capacity of 1mg 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 <1g 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 | Midi spin column |
| Purification technology | Silica technology, DNA filtration technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Plant leaves, roots, stems, fruits, fungi, etc. |
| Sample amount | 0.5-1 g |
| Elution volume | ≥400μl |
| Time per run | ≤40 minutes |
| Liquid carrying volume per column | 4ml |
| Binding yield of column | 1mg |
Principle
The Kit isolates total RNA from up to 1g plant tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 40 min. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanol is added to the flow-through, and the sample is applied to a RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 400µl 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 – medium isolation of several samples can be completed in 40 minutes
- 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 | R415202 | D415203 |
| Purification Times | 20 Preps | 100 Preps |
| HiPure DNA Midi Column II | 20 | 100 |
| HiPure RNA Midi Columns II | 20 | 100 |
| 15ml Collection Tubes | 40 | 200 |
| Buffer RLC | 120 ml | 550 ml |
| Buffer PRC1 | 120 ml | 550 ml |
| Buffer RW1 | 70 ml | 400 ml |
| Buffer RW2* | 20 ml | 100 ml |
| RNase Free Water | 30 ml | 120 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 in the Buffer RLC/PRC1. Dissolve by warming buffer to 37°C.
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