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 (not include miRNA) 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:≤1 x 107 Animal tissue sample: 1-20 mg Plant tissue: 50-150 mg |
Yield | 2-100μg |
Elution volume | ≥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.
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