Introduction
This kit provides fast purification of high-quality RNA from cells, 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 isopropanol precipitation. RNA purified using the HiPure 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 20mg tissue, 150mg plant, 5 x 106 cell using two columns (gDNA removed column) |
| Applications | RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing |
| Purification method | Mini spin column |
| Purification technology | Silica technology, DNA filtration technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Animal soft tissue, cultured cells, lymphocytes, simple plant tissue |
| Sample amount |
Cells:<span "="" style="outline: 0px; box-sizing: border-box; font-size: 9pt;">≤1 x 107 Animal tissue sample: 1-20 mg Plant tissue: 50-150 mg |
| Yield | 2-100μg |
| Elution volume | Variable to 50μl |
| Time per run | ≤25 minutes（1-24 samples） |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
Principle
The Kit isolates total RNA from up to 107 cells or 20 mg tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 mins. 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 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
Kit Contents
| Contents | R411102 | D411103 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure DNA Mini Columns |
50 |
250 |
| HiPure RNA Mini Columns | 50 | 250 |
| 2ml Collection Tubes | 100 | 2 x 250 |
| Buffer RLC |
50 ml |
200 ml |
| Buffer RW1 | 50 ml | 200 ml |
| Buffer RW2* |
12 ml |
2 x 50 ml |
| RNase Free Water |
10 ml |
30 ml |
Storage and Stability
HiPureKit can be stored dry at room temperature (15-25°C) and are stable for at least18 months under these conditions. During shipment, crystals or precipitationmay form in the Buffer RLC. Dissolve by warming buffer to 37°C.